Isoindoline compounds and methods of their use

ABSTRACT

Provided herein are isoindoline compounds, pharmaceutical compositions comprising one or more of such compounds, and methods of their use for treating, preventing, or managing various diseases.

This application is a continuation application of U.S. application Ser.No. 13/359,369, filed Jan. 26, 2012 now U.S. Pat. No. 8,222,249, nowallowed, which is a continuation application of U.S. application Ser.No. 12/608,953, filed Oct. 29, 2009, now U.S. Pat. No. 8,129,375, whichclaims priority to U.S. Provisional Application No. 61/109,475, filedOct. 29, 2008, the entireties of which are incorporated herein byreference.

1. FIELD

Provided herein are isoindoline compounds, pharmaceutical compositionscomprising one or more of such compounds, and methods of their use fortreating, preventing, or managing various diseases.

2. BACKGROUND 2.1 Pathobiology of Cancer and Other Diseases

Cancer is characterized primarily by an increase in the number ofabnormal cells derived from a given normal tissue, invasion of adjacenttissues by these abnormal cells, or lymphatic or blood-borne spread ofmalignant cells to regional lymph nodes and to distant sites(metastasis). Clinical data and molecular biologic studies indicate thatcancer is a multistep process that begins with minor preneoplasticchanges, which may under certain conditions progress to neoplasia. Theneoplastic lesion may evolve clonally and develop an increasing capacityfor invasion, growth, metastasis, and heterogeneity, especially underconditions in which the neoplastic cells escape the host's immunesurveillance. Roitt et al., Immunology 17.1-17.12 (3rd ed., Mosby, St.Louis, Mo., 1993).

There are an enormous variety of cancers which are described in detailin the medical literature. Examples include cancer of the lung, colon,rectum, prostate, breast, brain, and intestine. The incidence of cancercontinues to climb as the general population ages, as new cancersdevelop, and as susceptible populations (e.g., people infected with AIDSor excessively exposed to sunlight) grow. However, options for thetreatment of cancer are limited. For example, in the case of bloodcancers (e.g., multiple myeloma), few treatment options are available,especially when conventional chemotherapy fails and bone-marrowtransplantation is not an option. A tremendous demand therefore existsfor new methods and compositions that can be used to treat patients withcancer.

Many types of cancers are associated with new blood vessel formation, aprocess known as angiogenesis. Several of the mechanisms involved intumor-induced angiogenesis have been elucidated. The most direct ofthese mechanisms is the secretion by the tumor cells of cytokines withangiogenic properties. Examples of these cytokines include acidic andbasic fibroblastic growth factor (α,β-FGF), angiogenin, vascularendothelial growth factor (VEGF), and TNF-α. Alternatively, tumor cellscan release angiogenic peptides through the production of proteases andthe subsequent breakdown of the extracellular matrix where somecytokines are stored (e.g., β-FGF). Angiogenesis can also be inducedindirectly through the recruitment of inflammatory cells (particularlymacrophages) and their subsequent release of angiogenic cytokines (e.g.,TNF-α, β-FGF).

A variety of other diseases and disorders are also associated with, orcharacterized by, undesired angiogenesis. For example, enhanced orunregulated angiogenesis has been implicated in a number of diseases andmedical conditions including, but not limited to, ocular neovasculardiseases, choroidal neovascular diseases, retina neovascular diseases,rubeosis (neovascularization of the angle), viral diseases, geneticdiseases, inflammatory diseases, allergic diseases, and autoimmunediseases. Examples of such diseases and conditions include, but are notlimited to: diabetic retinopathy, retinopathy of prematurity, cornealgraft rejection, neovascular glaucoma, retrolental fibroplasia,arthritis, and proliferative vitreoretinopathy.

Accordingly, compounds that can control angiogenesis or inhibit theproduction of certain cytokines, including TNF-α, may be useful in thetreatment and prevention of various diseases and conditions.

2.2 Methods of Treating Cancer

Current cancer therapy may involve surgery, chemotherapy, hormonaltherapy and/or radiation treatment to eradicate neoplastic cells in apatient (see, e.g., Stockdale, 1998, Medicine, vol. 3, Rubenstein andFederman, eds., Chapter 12, Section IV). Recently, cancer therapy couldalso involve biological therapy or immunotherapy. All of theseapproaches pose significant drawbacks for the patient. Surgery, forexample, may be contraindicated due to the health of a patient or may beunacceptable to the patient. Additionally, surgery may not completelyremove neoplastic tissue. Radiation therapy is only effective when theneoplastic tissue exhibits a higher sensitivity to radiation than normaltissue. Radiation therapy can also often elicit serious side effects.Hormonal therapy is rarely given as a single agent. Although hormonaltherapy can be effective, it is often used to prevent or delayrecurrence of cancer after other treatments have removed the majority ofcancer cells. Biological therapies and immunotherapies are limited innumber and may produce side effects such as rashes or swellings,flu-like symptoms, including fever, chills and fatigue, digestive tractproblems, or allergic reactions.

With respect to chemotherapy, there is a variety of chemotherapeuticagents available for treatment of cancer. A majority of cancerchemotherapeutics act by inhibiting DNA synthesis, either directly orindirectly by inhibiting the biosynthesis of deoxyribonucleotidetriphosphate precursors, to prevent DNA replication and concomitant celldivision. Gilman et al., Goodman and Gilman's: The Pharmacological Basisof Therapeutics, Tenth Ed. (McGraw Hill, New York).

Despite availability of a variety of chemotherapeutic agents,chemotherapy has many drawbacks. Stockdale, Medicine, vol. 3, Rubensteinand Federman, eds., Chapter. 12, sect. 10, 1998. Almost allchemotherapeutic agents are toxic, and chemotherapy causes significantand often dangerous side effects including severe nausea, bone marrowdepression, and immunosuppression. Additionally, even withadministration of combinations of chemotherapeutic agents, many tumorcells are resistant or develop resistance to the chemotherapeuticagents. In fact, those cells resistant to the particularchemotherapeutic agents used in the treatment protocol often prove to beresistant to other drugs, even if those agents act by differentmechanism from those of the drugs used in the specific treatment. Thisphenomenon is referred to as pleiotropic drug or multidrug resistance.Because of the drug resistance, many cancers prove or become refractoryto standard chemotherapeutic treatment protocols.

Other diseases or conditions associated with, or characterized by,undesired angiogenesis are also difficult to treat. However, somecompounds such as protamine, heparin and steroids have been proposed tobe useful in the treatment of certain specific diseases. Taylor et al.,Nature 297:307 (1982); Folkman et al., Science 221:719 (1983); and U.S.Pat. Nos. 5,001,116 and 4,994,443.

Still, there is a significant need for effective methods of treating,preventing, and managing cancer and other diseases and conditions,particularly for diseases that are refractory to standard treatments,such as surgery, radiation therapy, chemotherapy and hormonal therapy,while reducing or avoiding the toxicities and/or side effects associatedwith the conventional therapies.

3. SUMMARY

Provided herein are isoindoline compounds, and pharmaceuticallyacceptable salts, solvates, prodrugs, or stereoisomers thereof.

Also provided herein are pharmaceutical compositions comprising acompound provided herein, e.g., a compound of Formula I, including asingle enantiomer, a mixture of enantiomers, or a mixture ofdiastereomers thereof; or a pharmaceutically acceptable salt, solvate,or prodrug thereof: in combination with one or more pharmaceuticallyacceptable carriers.

Further provided herein are methods of treating, preventing, or managingvarious diseases in a subject, which comprise administering to thesubject a therapeutically effective amount of a compound providedherein, e.g., a compound of Formula I, including a single enantiomer, amixture of enantiomers, or a mixture of diastereomers thereof, or apharmaceutically acceptable salt, solvate, or prodrug thereof.

4. DETAILED DESCRIPTION 4.1 Definitions

To facilitate understanding of the disclosure set forth herein, a numberof terms are defined below.

Generally, the nomenclature used herein and the laboratory procedures inorganic chemistry, medicinal chemistry, and pharmacology describedherein are those well known and commonly employed in the art. Unlessdefined otherwise, all technical and scientific terms used hereingenerally have the same meaning as commonly understood by one ofordinary skill in the art to which this disclosure belongs.

The term “subject” refers to an animal, including, but not limited to, aprimate (e.g., human), cow, sheep, goat, horse, dog, cat, rabbit, rat,or mouse. The terms “subject” and “patient” are used interchangeablyherein in reference, for example, to a mammalian subject, such as ahuman subject.

The terms “treat,” “treating,” and “treatment” refer to the eradicationor amelioration of a disease or disorder, or of one or more symptomsassociated with the disease or disorder. In general, treatment occursafter the onset of the disease or disorder. In certain embodiments, theterms refer to minimizing the spread or worsening of the disease ordisorder resulting from the administration of one or more prophylacticor therapeutic agents to a subject with such a disease or disorder.

The terms “prevent,” “preventing,” and “prevention” refer to theprevention of the onset, recurrence or spread of a disease or disorder,or of one or more symptoms thereof. In general, prevention occurs priorto the onset of the disease or disorder.

The terms “manage,” “managing,” and “management” refer to preventing orslowing the progression, spread or worsening of a disease or disorder,or of one or more symptoms thereof. Sometimes, the beneficial effectsthat a subject derives from a prophylactic or therapeutic agent do notresult in a cure of the disease or disorder.

The term “therapeutically effective amount” are meant to include theamount of a compound that, when administered, is sufficient to preventdevelopment of or alleviate to some extent, one or more of the symptomsof the disorder, disease, or condition being treated. The term“therapeutically effective amount” also refers to the amount of acompound that is sufficient to elicit the biological or medical responseof a cell, tissue, system, animal, or human, which is being sought by aresearcher, veterinarian, medical doctor, or clinician.

The term “IC₅₀” refers an amount, concentration, or dosage of a compoundthat is required for 50% inhibition of a maximal response in an assaythat measures such response.

The term “pharmaceutically acceptable carrier,” “pharmaceuticallyacceptable excipient,” “physiologically acceptable carrier,” or“physiologically acceptable excipient” refers to apharmaceutically-acceptable material, composition, or vehicle, such as aliquid or solid filler, diluent, excipient, solvent, or encapsulatingmaterial. In one embodiment, each component is “pharmaceuticallyacceptable” in the sense of being compatible with the other ingredientsof a pharmaceutical formulation, and suitable for use in contact withthe tissue or organ of humans and animals without excessive toxicity,irritation, allergic response, immunogenicity, or other problems orcomplications, commensurate with a reasonable benefit/risk ratio. See,Remington: The Science and Practice of Pharmacy, 21st Edition,Lippincott Williams & Wilkins: Philadelphia, Pa., 2005; Handbook ofPharmaceutical Excipients, 5th Edition, Rowe et al., Eds., ThePharmaceutical Press and the American Pharmaceutical Association: 2005;and Handbook of Pharmaceutical Additives, 3rd Edition, Ash and Ash Eds.,Gower Publishing Company: 2007; Pharmaceutical Preformulation andFormulation, Gibson Ed., CRC Press LLC: Boca Raton, Fla., 2004.

The term “about” or “approximately” means an acceptable error for aparticular value as determined by one of ordinary skill in the art,which depends in part on how the value is measured or determined. Incertain embodiments, the term “about” or “approximately” means within 1,2, 3, or 4 standard deviations. In certain embodiments, the term “about”or “approximately” means within 50%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%,4%, 3%, 2%, 1%, 0.5%, or 0.05% of a given value or range.

The terms “active ingredient” and “active substance” refer to acompound, which is administered, alone or in combination with one ormore pharmaceutically acceptable excipients, to a subject for treating,preventing, or ameliorating one or more symptoms of a condition,disorder, or disease. As used herein, “active ingredient” and “activesubstance” may be an optically active isomer of a compound describedherein.

The terms “drug,” “therapeutic agent,” and “chemotherapeutic agent”refer to a compound, or a pharmaceutical composition thereof, which isadministered to a subject for treating, preventing, or ameliorating oneor more symptoms of a condition, disorder, or disease.

The term “alkyl” refers to a linear or branched saturated monovalenthydrocarbon radical, wherein the alkyl may optionally be substitutedwith one or more substituents. The term “alkyl” also encompasses bothlinear and branched alkyl, unless otherwise specified. In certainembodiments, the alkyl is a linear saturated monovalent hydrocarbonradical that has 1 to 20 (C₁₋₂₀), 1 to 15 (C₁₋₁₅), 1 to 12 (C₁₋₁₂), 1 to10 (C₁₋₁₀), or 1 to 6 (C₁₋₆) carbon atoms, or branched saturatedmonovalent hydrocarbon radical of 3 to 20 (C₃₋₂₀), 3 to 15 (C₃₋₁₅), 3 to12 (C₃₋₁₂), 3 to 10 (C₃₋₁₀), or 3 to 6 (C₁₋₆) carbon atoms. As usedherein, linear C₁₋₆ and branched C₃₋₆ alkyl groups are also referred as“lower alkyl.” Examples of alkyl groups include, but are not limited to,methyl, ethyl, propyl (including all isomeric forms), n-propyl,isopropyl, butyl (including all isomeric forms), n-butyl, isobutyl,t-butyl, pentyl (including all isomeric forms), and hexyl (including allisomeric forms). For example, C₁₋₆ alkyl refers to a linear saturatedmonovalent hydrocarbon radical of 1 to 6 carbon atoms or a branchedsaturated monovalent hydrocarbon radical of 3 to 6 carbon atoms.

The term “alkenyl” refers to a linear or branched monovalent hydrocarbonradical, which contains one or more, in one embodiment, one to five,carbon-carbon double bonds. The alkenyl may be optionally substitutedone or more substituents. The term “alkenyl” also embraces radicalshaving “cis” and “trans” configurations, or alternatively, “E” and “Z”configurations, as appreciated by those of ordinary skill in the art. Asused herein, the term “alkenyl” encompasses both linear and branchedalkenyl, unless otherwise specified. For example, C₂₋₆ alkenyl refers toa linear unsaturated monovalent hydrocarbon radical of 2 to 6 carbonatoms or a branched unsaturated monovalent hydrocarbon radical of 3 to 6carbon atoms. In certain embodiments, the alkenyl is a linear monovalenthydrocarbon radical of 2 to 20 (C₂₋₂₀), 2 to 15 (C₂₋₁₅), 2 to 12(C₂₋₁₂), 2 to 10 (C₂₋₁₀), or 2 to 6 (C₂₋₆) carbon atoms, or a branchedmonovalent hydrocarbon radical of 3 to 20 (C₁₋₂₀), 3 to 15 (C₃₋₁₅), 3 to12 (C₃₋₁₂), 3 to 10 (C₃₋₁₀), or 3 to 6 (C₃₋₆) carbon atoms. Examples ofalkenyl groups include, but are not limited to, ethenyl, propen-1-yl,propen-2-yl, allyl, butenyl, and 4-methylbutenyl.

The term “alkynyl” refers to a linear or branched monovalent hydrocarbonradical, which contains one or more, in one embodiment, one to five,carbon-carbon triple bonds. The alkynyl may be optionally substitutedone or more substituents. The term “alkynyl” also encompasses bothlinear and branched alkynyl, unless otherwise specified. In certainembodiments, the alkynyl is a linear monovalent hydrocarbon radical of 2to 20 (C₂₋₂₀), 2 to 15 (C₂₋₁₅), 2 to 12 (C₂₋₁₂), 2 to 10 (C₂₋₁₀), or 2to 6 (C₂₋₆) carbon atoms, or a branched monovalent hydrocarbon radicalof 3 to 20 (C₃₋₁₀), 3 to 15 (C₃₋₁₅), 3 to 12 (C₃₋₁₂), 3 to 10 (C₃₋₁₀),or 3 to 6 (C₃₋₆) carbon atoms. Examples of alkynyl groups include, butare not limited to, ethynyl (—C≡CH) and propargyl (—CH₂C≡CH). Forexample, C₂₋₆ alkynyl refers to a linear unsaturated monovalenthydrocarbon radical of 2 to 6 carbon atoms or a branched unsaturatedmonovalent hydrocarbon radical of 3 to 6 carbon atoms.

The term “cycloalkyl” refers to a cyclic saturated bridged and/ornon-bridged monovalent hydrocarbon radical, which may be optionallysubstituted one or more substituents. In certain embodiments, thecycloalkyl has from 3 to 20 (C₃₋₁₀), from 3 to 15 (C₃₋₁₅), from 3 to 12(C₃₋₁₂), from 3 to 10 (C₃₋₁₀), or from 3 to 7 (C₃₋₇) carbon atoms.Examples of cycloalkyl groups include, but are not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,decalinyl, and adamantyl.

The term “aryl” refers to a monocyclic aromatic group and/or multicyclicmonovalent aromatic group that contain at least one aromatic hydrocarbonring. In certain embodiments, the aryl has from 6 to 20 (C₆₋₂₀), from 6to 15 (C₆₋₁₅), or from 6 to 10 (C₆₋₁₀) ring atoms. Examples of arylgroups include, but are not limited to, phenyl, naphthyl, fluorenyl,azulenyl, anthryl, phenanthryl, pyrenyl, biphenyl, and terphenyl. Arylalso refers to bicyclic or tricyclic carbon rings, where one of therings is aromatic and the others of which may be saturated, partiallyunsaturated, or aromatic, for example, dihydronaphthyl, indenyl,indanyl, or tetrahydronaphthyl (tetralinyl). In certain embodiments,aryl may also be optionally substituted with one or more substituents.

The term “aralkyl” or “aryl-alkyl” refers to a monovalent alkyl groupsubstituted with aryl. In certain embodiments, both alkyl and aryl maybe optionally substituted with one or more substituents.

The term “heteroaryl” refers to a monocyclic aromatic group and/ormulticyclic aromatic group that contain at least one aromatic ring,wherein at least one aromatic ring contains one or more heteroatomsindependently selected from O, S, and N. Each ring of a heteroaryl groupcan contain one or two O atoms, one or two S atoms, and/or one to four Natoms, provided that the total number of heteroatoms in each ring isfour or less and each ring contains at least one carbon atom. Theheteroaryl may be attached to the main structure at any heteroatom orcarbon atom which results in the creation of a stable compound. Incertain embodiments, the heteroaryl has from 5 to 20, from 5 to 15, orfrom 5 to 10 ring atoms. Examples of monocyclic heteroaryl groupsinclude, but are not limited to, pyrrolyl, pyrazolyl, pyrazolinyl,imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl,furanyl, thienyl, oxadiazolyl, pyridyl, pyrazinyl, pyrimidinyl,pyridazinyl, and triazinyl. Examples of bicyclic heteroaryl groupsinclude, but are not limited to, indolyl, benzothiazolyl, benzoxazolyl,benzothienyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl,benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl,isobenzofuranyl, chromonyl, coumarinyl, cinnolinyl, quinoxalinyl,indazolyl, purinyl, pyrrolopyridinyl, furopyridinyl, thienopyridinyl,dihydroisoindolyl, and tetrahydroquinolinyl. Examples of tricyclicheteroaryl groups include, but are not limited to, carbazolyl,benzindolyl, phenanthrollinyl, acridinyl, phenanthridinyl, andxanthenyl. In certain embodiments, heteroaryl may also be optionallysubstituted with one or more substituents.

The term “heterocyclyl” or “heterocyclic” refers to a monocyclicnon-aromatic ring system and/or multicyclic ring system that contains atleast one non-aromatic ring, wherein one or more of the non-aromaticring atoms are heteroatoms independently selected from O, S, or N; andthe remaining ring atoms are carbon atoms. In certain embodiments, theheterocyclyl or heterocyclic group has from 3 to 20, from 3 to 15, from3 to 10, from 3 to 8, from 4 to 7, or from 5 to 6 ring atoms. In certainembodiments, the heterocyclyl is a monocyclic, bicyclic, tricyclic, ortetracyclic ring system, which may includes a fused or bridged ringsystem, and in which the nitrogen or sulfur atoms may be optionallyoxidized, the nitrogen atoms may be optionally quaternized, and somerings may be partially or fully saturated, or aromatic. The heterocyclylmay be attached to the main structure at any heteroatom or carbon atomwhich results in the creation of a stable compound. Examples of suchheterocyclic radicals include, but are not limited to, acridinyl,azepinyl, benzimidazolyl, benzindolyl, benzoisoxazolyl, benzisoxazinyl,benzodioxanyl, benzodioxolyl, benzofuranonyl, benzofuranyl,benzonaphthofuranyl, benzopyranonyl, benzopyranyl,benzotetrahydrofuranyl, benzotetrahydrothienyl, benzothiadiazolyl,benzothiazolyl, benzothiophenyl, benzotriazolyl, benzothiopyranyl,benzoxazinyl, benzoxazolyl, benzothiazolyl, β-carbolinyl, carbazolyl,chromanyl, chromonyl, cinnolinyl, coumarinyl, decahydroisoquinolinyl,dibenzofuranyl, dihydrobenzisothiazinyl, dihydrobenzisoxazinyl,dihydrofuryl, dihydropyranyl, dioxolanyl, dihydropyrazinyl,dihydropyridinyl, dihydropyrazolyl, dihydropyrimidinyl, dihydropyrrolyl,dioxolanyl, 1,4-dithianyl, furanonyl, furanyl, imidazolidinyl,imidazolinyl, imidazolyl, imidazopyridinyl, imidazothiazolyl, indazolyl,indolinyl, indolizinyl, indolyl, isobenzotetrahydrofuranyl,isobenzotetrahydrothienyl, isobenzothienyl, isochromanyl, isocoumarinyl,isoindolinyl, isoindolyl, isoquinolinyl, isothiazolidinyl, isothiazolyl,isoxazolidinyl, isoxazolyl, morpholinyl, naphthyridinyl,octahydroindolyl, octahydroisoindolyl, oxadiazolyl, oxazolidinonyl,oxazolidinyl, oxazolopyridinyl, oxazolyl, oxiranyl, perimidinyl,phenanthridinyl, phenathrolinyl, phenarsazinyl, phenazinyl,phenothiazinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl,4-piperidonyl, pteridinyl, purinyl, pyrazinyl, pyrazolidinyl, pyrazolyl,pyridazinyl, pyridinyl, pyridopyridinyl, pyrimidinyl, pyrrolidinyl,pyrrolinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl,quinuclidinyl, tetrahydrofuryl, tetrahydrofuranyl,tertahydroisoquinolinyl, tetrahydropyranyl, tetrahydrothienyl,tetrazolyl, thiadiazolopyrimidinyl, thiadiazolyl, thiamorpholinyl,thiazolidinyl, thiazolyl, thienyl, triazinyl, triazolyl, and1,3,5-trithianyl. In certain embodiments, heterocyclic may also beoptionally substituted with one or more substituents.

The term “halogen”, “halide” or “halo” refers to fluorine, chlorine,bromine, and/or iodine.

The terms “optically active” and “enantiomerically active” refer to acollection of molecules, which has an enantiomeric excess of no lessthan about 50%, no less than about 70%, no less than about 80%, no lessthan about 90%, no less than about 91%, no less than about 92%, no lessthan about 93%, no less than about 94%, no less than about 95%, no lessthan about 96%, no less than about 97%, no less than about 98%, no lessthan about 99%, no less than about 99.5%, or no less than about 99.8%.

In describing an optically active compound, the prefixes R and S areused to denote the absolute configuration of the molecule about itschiral center(s). The (+) and (−) are used to denote the opticalrotation of the compound, that is, the direction in which a plane ofpolarized light is rotated by the optically active compound. The (−)prefix indicates that the compound is levorotatory, that is, thecompound rotates the plane of polarized light to the left orcounterclockwise. The (+) prefix indicates that the compound isdextrorotatory, that is, the compound rotates the plane of polarizedlight to the right or clockwise. However, the sign of optical rotation,(+) and (−), is not related to the absolute configuration of themolecule. R and S.

The term “solvate” refers to a compound provided herein or a saltthereof, which further includes a stoichiometric or non-stoichiometricamount of solvent bound by non-covalent intermolecular forces. Where thesolvent is water, the solvate is a hydrate.

4.2 Compounds

Provided herein are isoindoline compounds, pharmaceutical compositionscomprising one or more of such compounds, and methods of their use fortreating, preventing, or managing various diseases.

In one embodiment, provided is a compound of Formula I:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomerthereof, wherein:

X is C(═O) or CH₂;

Y is O, cyanamido (N—≡N), or amido (NH);

m is an integer of 0, 1, 2, or 3;

R¹ is hydrogen or C₁₋₆ alkyl;

R² is hydrogen, —NO₂, C₁₋₁₀ alkyl, C₀₋₆ alkyl-(5 to 10 memberedheteroaryl), C₀₋₆ alkyl-(5 to 6 membered heterocyclyl), C₀₋₆ alkyl-OH,C₀₋₄ alkyl-NH₂, —NHCO—C₁₋₆ alkyl, —OR²¹, or —(CH₂—Z)₀₋₂-(5 to 10membered heteroaryl), where each heteroaryl and heterocyclyl isoptionally substituted with one or more C₁₋₆ alkyl;

R³ is hydrogen, halogen, —NO₂, C₀₋₆ alkyl-(5 to 10 membered heteroaryl),C₀₋₆alkyl-(5 to 6 membered heterocyclyl), C₀₋₆alkyl-OH, C₀₋₄ alkyl-NH,—NHCO—C₁₋₆ alkyl, —OR²¹, or —(CH₂—Z)₀₋₂-(5 to 10 membered heteroaryl),where each heteroaryl and heterocyclyl is optionally substituted withone or more C₁₋₆ alkyl;

R²¹ is C₆₋₁₀ aryl, 5 to 10 membered heteroaryl, 5 to 6 memberedheterocyclyl, or —CO(CH₂)₀₋₂R²², wherein the aryl, heteroaryl, andheterocyclyl are each optionally substituted with one or more C₁₋₆alkyl;

R²² is —NH, or 5 to 6 membered heterocyclyl; and

Z is CH₂, NH, or O;

with the proviso that when R¹ is hydrogen, then R² is not hydrogen orC₁₋₁₀ alkyl;

with the proviso that when Y is O, then R³ is not halogen; and

with the proviso that when Y is O and R³ is halogen, then R² is C_(o-6)alkyl-(5-6 membered heterocyclyl).

In certain embodiments, X is CH₂. In certain embodiments, X is C(═O).

In certain embodiments, Y is O. In certain embodiments, Y is cyanamido.In certain embodiments, Y is amido.

In certain embodiments, Z is CH₂. In certain embodiments, Z is NH. Incertain embodiments, Z is O.

In certain embodiments, m is 0. In certain embodiments, m is 1. Incertain embodiments, m is 2. In certain embodiments, m is 3.

In certain embodiments, R¹ is hydrogen. In certain embodiments, R¹ isC₁₋₆ alkyl, optionally substituted with one, two, or three substituentsQ as described herein. In certain embodiments, R¹ is methyl.

In certain embodiments, R² is hydrogen. In certain embodiments, R² ishalogen. In certain embodiments. R² is nitro. In certain embodiments, R²is C₁₋₁₀ alkyl. In certain embodiments, R² is C₀₋₆ alkyl-(5 to 10membered heteroaryl), where the heteroaryl is optionally substitutedwith one or more C₁₋₆ alkyl. In certain embodiments, R² is C₀₋₆ alkyl-(5to 6 membered heterocyclyl), where the heterocyclyl is optionallysubstituted with one or more C₁₋₆ alkyl. In certain embodiments, R² isC₀₋₆ alkyl-OH. In certain embodiments, R² is C₀₋₄ alkyl-NH₂. In certainembodiments, R² is —NHCO— C₁₋₆ alkyl. In certain embodiments, R² is—OR²¹, wherein R²¹ is as described herein. In certain embodiments, R² isor —(CH₂—Y)₀₋₂-(5 to 10 membered heteroaryl), where the heteroaryl isoptionally substituted with one or more C₁₋₆ alkyl. In certainembodiments, R² is hydrogen, amino, acetamido, hydroxy, nitro,aminomethyl, hydroxymethyl, 2-methyl-1H-imidazol-1-yl,3-methyl-1,2,4-oxadiazol-5-yl, 4-methylpiperazin-1-yl)methyl,2-methyl-2H-pyrazol-3-yl, 1-methyl-1H-pyrazol-3-yl,2-methylthiazol-4-yl, 4-methyl-4H-1,2,4-triazol-3-yl, morpholinomethyl,(pyridin-4-yl)methyl, (pyridin-4-yloxy)methyl, pheoxy, pyridin-2-yloxy,piperidin-4-yloxy, 2-aminoacetoxy, or 2-piperazin-1-ylacetoxy.

In certain embodiments, R³ is hydrogen. In certain embodiments, R³ isnitro. In certain embodiments. R³ is C_(o-6) alkyl-(5 to 10 memberedheteroaryl), where the heteroaryl is optionally substituted with one ormore C₁₋₆ alkyl. In certain embodiments, R³ is C₀₋₆ alkyl-(5 to 6membered heterocyclyl), where the heterocyclyl is optionally substitutedwith one or more C₁₋₆ alkyl. In certain embodiments, R³ is C₀₋₆alkyl-OH. In certain embodiments, R³ is C₀₋₄ alkyl-NH). In certainembodiments, R³ is —NHCO—C₁₋₆ alkyl. In certain embodiments, R³ is—OR²¹, wherein R²¹ is as described herein. In certain embodiments, R³ isor —(CH₂—Y)₀₋₂-(5 to 10 membered heteroaryl), where the heteroaryl isoptionally substituted with one or more C₁₋₆ alkyl. In certainembodiments, R³ is hydrogen, amino, acetamido, hydroxy, nitro, methyl,aminomethyl, hydroxymethyl, imidazol-1-yl,3-methyl-1,2,4-oxadiazol-5-yl, 4-methylpiperazin-1-yl)methyl,2-methyl-2H-pyrazol-3-yl, 1-methyl-1H-pyrazol-3-yl,2-methylthiazol-4-yl, 4-methyl-4H-1,2,4-triazol-3-yl, morpholinomethyl,(pyridin-4-yl)methyl, (pyridin-4-yloxy)methyl, pheoxy, pyridin-2-yloxy,piperidin-4-yloxy, 2-aminoacetoxy, or 2-piperazin-1-ylacetoxy.

In one embodiment, the compound is:

or a pharmaceutically acceptable salt, solvate, prodrug, andstereoisomer thereof.

In another embodiment, provided herein is a compound of Formula II:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomerthereof, wherein:

X is C(═O) or CH₂;

m is an integer of 0, 1, 2, or 3;

R⁴ is C₃₋₁₀ cycloalkyl, 5 to 10 membered heterocyclyl, 5 to 10 memberedheteroaryl, or C₀₋₄ alkyl-NR⁴¹R⁴²; wherein the cycloalkyl, heterocyclyl,and heteroaryl are each optionally substituted with one or more halogen,C₁₋₆ alkyl, —CO—NR⁴³R⁴⁴, —COOR⁴⁵, or C₀₋₄ alkyl-C₆₋₁₀ aryl, wherein thearyl itself may be optionally substituted with one or more halogen; and

R⁴¹, R⁴², R⁴³, R⁴⁴ and R⁴⁵ are each independently hydrogen or C₁₋₆alkyl.

In certain embodiments, X is CH₂. In certain embodiments, X is C(═O).

In certain embodiments, m is 0. In certain embodiments, m is 1. Incertain embodiments, m is 2. In certain embodiments, m is 3.

In certain embodiments, R⁴ is C₃₋₁₀ cycloalkyl, optionally substitutedwith one or more (C₁₋₆) alkyl or C₀₋₄ alkyl-C₆₋₁₀ aryl. In certainembodiments, R⁴ is 5 to 6 membered heterocyclyl, optionally substitutedwith one or more (C₁₋₆) alkyl or C₀₋₄ alkyl-C₆₋₁₀ aryl. In certainembodiments, R⁴ is C₀₋₄ alkyl-NR⁴¹R⁴², wherein R⁴¹ and R⁴² are eachdescribed herein.

In certain embodiments, R⁴ is 3-(N,N-diethylamino)propyl,4-acetamidophenyl, 3-(2-aminoacetoxy)-4-methylphenyl,3-aminomethyl-4-methylphenyl, 2-aminomethyl-5-methylphenyl,3-aminophenyl, 3-amino-4-methylphenyl, 3-chloro-4-methylphenyl,4-hydroxymethylphenyl, 3-hydroxy-4-methylphenyl,3-(2-methyl-1H-imidazol-1-yl)phenyl, 4-methyl-3-nitrophenyl,3-(3-methyl-1,2,4-oxadiazol-5-yl)phenyl,4-methyl-3-(2-piperazin-1-ylacetoxy)-phenyl,3-((4-methylpiperazin-1-yl)methyl)phenyl,3-(1-methyl-1H-pyrazol-3-yl)phenyl, 3-(2-methyl-2H-pyrazol-3-yl)phenyl,3-(2-methylthiazol-4-yl)phenyl,4-(4-methyl-4H-1,2,4-triazol-3-yl)phenyl, 3-(morpholinomethyl)phenyl,4-(morpholinomethyl)phenyl, 4-nitrophenyl, phenyl,3-(piperidin-4-yloxy)phenyl, 4-(pyridin-4-yl)methylphenyl,4-((pyridin-4-yloxy)methyl)phenyl, 3-(pyridin-2-yloxy)phenyl,3-phenoxyphenyl, 4-tert-butylcyclohexyl, cis-4-tert-butylcyclohexyl,trans-4-tert-butylcyclohexyl, 4-methylcyclohexyl,cis-4-methylcyclohexyl, trans-4-methylcyclohexyl,1-benzylpiperidin-4-yl, 4-methyltetrahydro-2H-pyran-4-yl,piperidin-4-yl, 4-phenylcyclohexyl, cis-4-phenylcyclohexyl, ortrans-4-phenylcyclohexyl.

In one embodiment, the compound is:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomerthereof.

In yet another embodiment, provided herein is a compound of Formula III:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomerthereof, wherein:

X is C(═O) or CH₂;

m is an integer of 0, 1, 2, or 3;

R⁵ and R⁶ are each independently: hydrogen, halo, C₁₋₆ alkyl, oxo, —NO₂,C₁₋₆ alkoxy, —Z—C₁₋₆ alkyl, C₀₋₆ alkyl-(5 to 10 membered heteroaryl),C₀₋₆ alkyl-(5 to 6 membered heterocyclyl), C₀₋₆ alkyl-OH, C₀₋₄alkyl-NH₂, —NHCO—C₁₋₆ alkyl, —OR²¹, or —(CH₂—Y)₀₋₂-(5 to 10 memberedheteroaryl),

wherein Z is S or SO₂;

wherein R²¹ is as defined above;

wherein each heteroaryl and heterocyclyl above is optionally substitutedwith one or more C₁₋₆ alkyl; and

wherein the alkyl or alkoxy above may be optionally substituted with oneor more: halogen; cyano; nitro; amino; C₁₋₆ alkylidenedioxy; C₁₋₆alkoxy,itself optionally substituted with one or more halogens; or C₁₋₆alkylthio, itself optionally substituted with one or more halogens;

R⁷ is —COR⁷¹ or —PO(OR⁷²)(OR⁷³);

R⁷¹ is C₁₋₁₀ alkyl, C₆₋₁₀ aryl, or 5 to 6 membered heterocyclyl; whereinthe alkyl, aryl, heterocyclyl may be optionally substituted with one ormore amino, C₁₋₆ alkylamino, di(C₁₋₆ alkyl)amino, or —COOR⁷⁴; and

R⁷², R⁷³, and R⁷⁴ are each independently hydrogen or C₁₋₁₀ alkyl.

In certain embodiments, X is CH₂. In certain embodiments, X is C(═O).

In certain embodiments, m is 0. In certain embodiments, m is 1. Incertain embodiments, m is 2. In certain embodiments, m is 3.

In certain embodiments, R⁵ is hydrogen. In certain embodiments, R⁵ ishalo. In certain embodiments, R⁵ is fluoro or chloro.

In certain embodiments, R⁶ is hydrogen. In certain embodiments, R⁶ ishalo. In certain embodiments, R⁶ is fluoro or chloro.

In certain embodiments, R⁷ is —COR⁴¹, wherein R⁴¹ is as describedherein. In certain embodiments, R⁷ is —PO(OR⁴²))(OR⁴³), wherein R⁴² andR⁴³ are each as described herein.

In one embodiment, the compound is:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomerthereof, wherein R⁵ and R⁶ are as defined above.

In yet another embodiment, provided herein is a compound of Formula IV:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomerthereof, wherein:

X is C(═O) or CH₂;

n is an integer of 0 or 1;

R⁸ is hydrogen or halo; and

R⁹ is hydrogen, amino, or 5 to 10 membered heteroaryl or heterocyclyl;

with the proviso that when m is 0, R⁹ is not hydrogen.

In certain embodiments, X is CH₂. In certain embodiments, X is C(═O).

In certain embodiments, n is 0. In certain embodiments, n is 1.

In certain embodiments, R⁸ is hydrogen. In certain embodiments, R⁸ ishalo. In certain embodiments, R⁸ is fluoro or chloro.

In certain embodiments, R⁹ is hydrogen. In certain embodiments, R⁹ isamino. In certain embodiments, R⁹ is 5 to 10 membered heteroaryl. Incertain embodiments, R⁹ is 5 to 10 membered heterocyclyl.

In one embodiment, the compound is:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomerthereof.

In yet another embodiment, provided herein is a compound of Formula V:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomerthereof, wherein:

X is C(═O) or CH₂;

m is an integer of 0, 1, 2, or 3;

R¹⁰ and R¹¹ are each independently hydrogen, halo, C₁₋₆ alkyl, or C₆₋₁₀aryloxy,

wherein the alkyl and aryl are each optionally substituted with one ormore halo.

In certain embodiments, X is CH₂. In certain embodiments, X is C(═O).

In certain embodiments, m is 0. In certain embodiments, m is 1. Incertain embodiments, m is 2. In certain embodiments, m is 3.

In certain embodiments, R¹⁰ is hydrogen. In certain embodiments, R¹¹ ishalo. In certain embodiments, R¹⁰ is fluoro or chloro. In certainembodiments, R¹⁰ is C₁₋₆ alkyl, optionally substituted with one or morehalo. In certain embodiments, R¹⁰ is C₆₋₁₀ aryloxy, optionallysubstituted with one or more halo.

In certain embodiments, R¹¹ is hydrogen. In certain embodiments, R¹¹ ishalo. In certain embodiments, R¹¹ is fluoro or chloro. In certainembodiments, R¹¹ is C₁₋₆ alkyl, optionally substituted with one or morehalo. In certain embodiments, R¹¹ is C₆₋₁₀ aryloxy, optionallysubstituted with one or more halo.

In one embodiment, the compound is:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomerthereof.

In yet another embodiment, provided herein is a compound of Formula VI:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomerthereof, wherein:X is CH, or C═Om and n are each independently 0 or 1;p is 0, 1, 2, or 3;R⁸¹ is 5 to 6 membered heterocyclyl, optionally substituted with C₁₋₆alkyl; andR⁸² is hydrogen or halogen.

In one embodiment, X is CH₂. In another embodiment, X is C═O.

In one embodiment, m is 0. In another embodiment, m is 1. In anotherembodiment, n is 0. In another embodiment, n is 1.

In one embodiment, p is 0. In another embodiment, p is 1. In anotherembodiment, p is 2. In another embodiment, p is 3.

In one embodiment, R⁸¹ is 5 membered heterocycle. In another embodiment,the 5 membered heterocycle is substituted with C₁₋₆ alkyl. In anotherembodiment, R⁸¹ is 6 membered heterocycle. In another embodiment, the 6membered heterocycle is substituted with C₁₋₆ alkyl.

In one embodiment, R⁸² is hydrogen. In another embodiment, R⁸² ishalogen.

In one embodiment, the compound is:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomerthereof.

In yet another embodiment, provided herein is a compound of thefollowing formula:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomerthereof.

The compounds provided herein are intended to encompass all possiblestereoisomers, unless a particular stereochemistry is specified. Wherethe compound provided herein contains an alkenyl or alkenylene group,the compound may exist as one or mixture of geometric cis/trans (or Z/E)isomers. Where structural isomers are interconvertible via a low energybarrier, the compound may exist as a single tautomer or a mixture oftautomers. This can take the form of proton tautomerism in the compoundthat contains, for example, an imino, keto, or oxime group; or so-calledvalence tautomerism in the compound that contain an aromatic moiety. Itfollows that a single compound may exhibit more than one type ofisomerism.

The compounds provided herein may be enantiomerically pure, such as asingle enantiomer or a single diastereomer, or be stereoisomericmixtures, such as a mixture of enantiomers, a racemic mixture, or adiastereomeric mixture. As such, one of skill in the art will recognizethat administration of a compound in its (R) form is equivalent, forcompounds that undergo epimerization in vivo, to administration of thecompound in its (S) form. Conventional techniques for thepreparation/isolation of individual enantiomers include synthesis from asuitable optically pure precursor, asymmetric synthesis from achiralstarting materials, or resolution of an enantiomeric mixture, forexample, chiral chromatography, recrystallization, resolution,diastereomeric salt formation, or derivatization into diastereomericadducts followed by separation.

When the compound provided herein contains an acidic or basic moiety, itmay also be provided as a pharmaceutically acceptable salt (See, Bergeet al., J. Pharm. Sci. 1977, 66, 1-19; and “Handbook of PharmaceuticalSalts, Properties, and Use,” Stahl and Wermuth, Ed.; Wiley-VCH and VHCA,Zurich, 2002).

Suitable acids for use in the preparation of pharmaceutically acceptablesalts include, but are not limited to, acetic acid, 2,2-dichloroaceticacid, acylated amino acids, adipic acid, alginic acid, ascorbic acid,L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoicacid, boric acid, (+)-camphoric acid, camphorsulfonic acid,(+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylicacid, cinnamic acid, citric acid, cyclamic acid, cyclohexanesulfamicacid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonicacid, 2-hydroxy-ethanesulfonic acid, formic acid, fumaric acid,galactaric acid, gentisic acid, glucoheptonic acid, D-gluconic acid,D-glucuronic acid, L-glutamic acid, α-oxoglutaric acid, glycolic acid,hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid,(+)-L-lactic acid, (±)-DL-lactic acid, lactobionic acid, lauric acid,maleic acid, (−)-L-malic acid, malonic acid, (±)-DL-mandelic acid,methanesulfonic acid, naphthalene-2-sulfonic acid,naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinicacid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid,pamoic acid, perchloric acid, phosphoric acid, L-pyroglutamic acid,saccharic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid,stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaricacid, thiocyanic acid, p-toluenesulfonic acid, undecylenic acid, andvaleric acid.

Suitable bases for use in the preparation of pharmaceutically acceptablesalts, including, but not limited to, inorganic bases, such as magnesiumhydroxide, calcium hydroxide, potassium hydroxide, zinc hydroxide, orsodium hydroxide; and organic bases, such as primary, secondary,tertiary, and quaternary, aliphatic and aromatic amines, includingL-arginine, benethamine, benzathine, choline, deanol, diethanolamine,diethylamine, dimethylamine, dipropylamine, diisopropylamine,2-(diethylamino)-ethanol, ethanolamine, ethylamine, ethylenediamine,isopropylamine, N-methyl-glucamine, hydrabamine, 1H-imidazole, L-lysine,morpholine, 4-(2-hydroxyethyl)-morpholine, methylamine, piperidine,piperazine, propylamine, pyrrolidine, 1-(2-hydroxyethyl)-pyrrolidine,pyridine, quinuclidine, quinoline, isoquinoline, secondary amines,triethanolamine, trimethylamine, triethylamine, N-methyl-D-glucamine,2-amino-2-(hydroxymethyl)-1,3-propanediol, and tromethamine.

The compound provided herein may also be provided as a prodrug, which isa functional derivative of the compound and is readily convertible intothe parent compound in vivo. Prodrugs are often useful because, in somesituations, they may be easier to administer than the parent compound.They may, for instance, be bioavailable by oral administration whereasthe parent compound is not. The prodrug may also have enhancedsolubility in pharmaceutical compositions over the parent compound. Aprodrug may be converted into the parent drug by various mechanisms,including enzymatic processes and metabolic hydrolysis. See Harper,Progress in Drug Research 1962, 4, 221-294; Morozowich et al. in “Designof Biopharmaceutical Properties through Prodrugs and Analogs,” RocheEd., APHA Acad. Pharm. Sci. 1977; “Bioreversible Carriers in Drug inDrug Design, Theory and Application,” Roche Ed., APHA Acad. Pharm. Sci.1987; “Design of Prodrugs,” Bundgaard, Elsevier, 1985; Wang et al.,Curr. Pharm. Design 1999, 5, 265-287; Pauletti et al., Adv. Drug.Delivery Rev. 1997, 27, 235-256; Mizen et al., Pharm. Biotech. 1998, 11,345-365; Gaignault et al., Pract. Med. Chem. 1996, 671-696; Asgharnejadin “Transport Processes in Pharmaceutical Systems,” Amidon et al., Ed.,Marcell Dekker, 185-218, 2000; Balant et al., Eur. J. Drug Metab.Pharmacokinet. 1990, 15, 143-53; Balimane and Sinko, Adv. Drug DeliveryRev. 1999, 39, 183-209; Browne. Clin. Neuropharmacol. 1997, 20, 1-12;Bundgaard, Arch. Pharm. Chem. 1979, 86, 1-39; Bundgaard, Controlled DrugDelivery 1987, 17, 179-96; Bundgaard, Adv. Drug Delivery Rev. 1992, 8,1-38; Fleisher et al., Adv. Drug Delivery Rev. 1996, 19, 115-130;Fleisher et al., Methods Enzymol. 1985, 112, 360-381; Farquhar et al.,J. Pharm. Sci. 1983, 72, 324-325; Freeman et al. J. Chem. Soc., Chem.Commun. 1991, 875-877; Friis and Bundgaard, Eur. J. Pharm. Sci. 1996, 4,49-59; Gangwar et al. Des. Biopharm. Prop. Prodrugs Analogs, 1977,409-421; Nathwani and Wood, Drugs 1993, 45, 866-94; Sinhababu andThakker, Adv. Drug Delivery Rev. 1996, 19, 241-273; Stella et al., Drugs1985, 29, 455-73; Tan et al., Adv. Drug Delivery Rev. 1999, 39, 117-151;Taylor, Adv. Drug Delivery Rev. 1996, 19, 131-148; Valentino andBorchardt, Drug Discovery Today 1997, 2, 148-155; Wiebe and Knaus, Adv.Drug Delivery Rev. 1999, 39, 63-80; and Waller et al., Br. J. Clin.Pharmac. 1989, 28, 497-507.

4.3 Methods of Treatment, Prevention and Management

In one embodiment, provided herein are methods of treating, preventing,and/or managing various diseases or disorders using a compound providedherein, including a single enantiomer, a mixture of enantiomers, or amixture of diastereomers thereof, or a pharmaceutically acceptable salt,solvate, or prodrug thereof. Without being limited by a particulartheory, compounds provided herein can control angiogenesis or inhibitthe production of certain cytokines including, but not limited to,TNF-α, IL-1β, IL-12, IL-18, GM-CSF, and/or IL-6. Without being limitedby a particular theory, compounds provided herein can stimulate theproduction of certain other cytokines including IL-10, and also act as acostimulatory signal for T cell activation, resulting in increasedproduction of cytokines such as, but not limited to, IL-12 and/or IFN-γ.In addition, compounds provided herein can enhance the effects of NKcells and antibody-mediated cellular cytotoxicity (ADCC). Further,compounds provided herein may be immunomodulatory and/or cytotoxic, andthus, may be useful as chemotherapeutic agents. Consequently, withoutbeing limited by a particular theory, some or all of suchcharacteristics possessed by the compounds provided herein may renderthem useful in treating, managing, and/or preventing various diseases ordisorders. In one embodiment, the compounds provided herein arecytotoxic.

The diseases or disorders treatable with the methods provided hereininclude, but are not limited to, cancer, disorders associated withangiogenesis, pain including Complex Regional Pain Syndrome (“CRPS”),Macular Degeneration (“MD”) and related syndromes, skin diseases,pulmonary disorders, asbestos-related disorders, parasitic diseases,immunodeficiency disorders, CNS disorders, CNS injury, atherosclerosisand related disorders, dysfunctional sleep and related disorders,hemoglobinopathy and related disorders (e.g., anemia), TNFα and othercytokines related disorders, and other various diseases and disorders.

Examples of cancer and precancerous conditions include, but are notlimited to, those described in U.S. Pat. Nos. 6,281,230 and 5,635,517 toMuller et al., in various U.S. patent publications to Zeldis, includingPubl. Nos. 2004/0220144A1, published Nov. 4, 2004 (Treatment ofMyelodysplastic Syndrome); 2004/0029832A1, published Feb. 12, 2004(Treatment of Various Types of Cancer); and 2004/0087546, published May6, 2004 (Treatment of Myeloproliferative Diseases). Examples alsoinclude those described in PCT/US04/14004, filed May 5, 2004. All ofthese references are incorporated herein in their entireties byreference.

Examples of cancer include, but are not limited to, cancers of the skin,such as melanoma; lymph node; breast; cervix; uterus; gastrointestinaltract; lung; ovary; prostate; colon; rectum; mouth; brain; head andneck; throat; testes; thyroid; kidney; pancreas; bone; spleen; liver;bladder; larynx; nasal passages; and AIDS-related cancers. The compoundsprovided here are useful for treating cancers of the blood and bonemarrow, such as multiple myeloma and acute and chronic leukemias, forexample, lymphoblastic, myelogenous, lymphocytic, and myelocyticleukemias. The compounds provided herein can be used for treating,preventing, or managing either primary or metastatic tumors.

Other examples of cancers include, but are not limited to, advancedmalignancy, amyloidosis, neuroblastoma, meningioma, hemangiopericytoma,multiple brain metastase, glioblastoma multiforms, glioblastoma, brainstem glioma, poor prognosis malignant brain tumor, malignant glioma,recurrent malignant glioma, anaplastic astrocytoma, anaplasticoligodendroglioma, neuroendocrine tumor, rectal adenocarcinoma, Dukes C& D colorectal cancer, unresectable colorectal carcinoma, metastatichepatocellular carcinoma, Kaposi's sarcoma, karotype acute myeloblasticleukemia, chronic lymphocytic leukemia (CLL), Hodgkin's lymphoma,non-Hodgkin's lymphoma, cutaneous T-Cell lymphoma, cutaneous B-Celllymphoma, diffuse large B-Cell lymphoma, low grade follicular lymphoma,metastatic melanoma (localized melanoma, including, but not limited to,ocular melanoma), malignant mesothelioma, malignant pleural effusionmesothelioma syndrome, peritoneal carcinoma, papillary serous carcinoma,gynecologic sarcoma, soft tissue sarcoma, scleroderma, cutaneousvasculitis, Langerhans cell histiocytosis, leiomyosarcoma,fibrodysplasia ossificans progressive, hormone refractory prostatecancer, resected high-risk soft tissue sarcoma, unrescectablehepatocellular carcinoma, Waldenstrom's macroglobulinemia, smolderingmyeloma, indolent myeloma, fallopian tube cancer, androgen independentprostate cancer, androgen dependent stage IV non-metastatic prostatecancer, hormone-insensitive prostate cancer, chemotherapy-insensitiveprostate cancer, papillary thyroid carcinoma, follicular thyroidcarcinoma, medullary thyroid carcinoma, and leiomyoma. In certainembodiments, the cancer is metastatic. In certain embodiments, thecancer is refractory or resistance to chemotherapy or radiation.

In another embodiment, provided herein are methods of treating,preventing or managing various forms of leukemias such as chroniclymphocytic leukemia, chronic myelocytic leukemia, acute lymphoblasticleukemia, acute myelogenous leukemia and acute myeloblastic leukemia,including leukemias that are relapsed, refractory or resistant, asdisclosed in U.S. Publ. No. 2006/0030594, published Feb. 9, 2006, whichis incorporated in its entirety by reference. The term “leukemia” refersmalignant neoplasms of the blood-forming tissues. The leukemia includes,but is not limited to, chronic lymphocytic leukemia, chronic myelocyticleukemia, acute lymphoblastic leukemia, acute myelogenous leukemia andacute myeloblastic leukemia. The leukemia can be relapsed, refractory orresistant to conventional therapy. The term “relapsed” refers to asituation where patients who have had a remission of leukemia aftertherapy have a return of leukemia cells in the marrow and a decrease innormal blood cells. The term “refractory or resistant” refers to acircumstance where patients, even after intensive treatment, haveresidual leukemia cells in their marrow.

In yet another embodiment, provided herein are methods of treating,preventing or managing various types of lymphomas, includingNon-Hodgkin's lymphoma (NHL). The term “lymphoma” refers a heterogenousgroup of neoplasms arising in the reticuloendothelial and lymphaticsystems. “NHL” refers to malignant monoclonal proliferation of lymphoidcells in sites of the immune system, including lymph nodes, bone marrow,spleen, liver and gastrointestinal tract. Examples of NHL include, butare not limited to, mantle cell lymphoma, MCL, lymphocytic lymphoma ofintermediate differentiation, intermediate lymphocytic lymphoma, ILL,diffuse poorly differentiated lymphocytic lymphoma, PDL, centrocyticlymphoma, diffuse small-cleaved cell lymphoma, DSCCL, follicularlymphoma, and any type of the mantle cell lymphomas that can be seenunder the microscope (nodular, diffuse, blastic and mentle zonelymphoma).

Examples of diseases and disorders associated with, or characterized by,undesired angiogenesis include, but are not limited to, inflammatorydiseases, autoimmune diseases, viral diseases, genetic diseases,allergic diseases, bacterial diseases, ocular neovascular diseases,choroidal neovascular diseases, retina neovascular diseases, andrubeosis (neovascularization of the angle). Specific examples of thediseases and disorders associated with, or characterized by, undesiredangiogenesis include, but are not limited to, endometriosis, Crohn'sdisease, heart failure, advanced heart failure, renal impairment,endotoxemia, toxic shock syndrome, osteoarthritis, retrovirusreplication, wasting, meningitis, silica-induced fibrosis,asbestos-induced fibrosis, veterinary disorder, malignancy-associatedhypercalcemia, stroke, circulatory shock, periodontitis, gingivitis,macrocytic anemia, refractory anemia, and 5q-deletion syndrome.

Examples of pain include, but are not limited to those described in U.S.patent Publ. No. 2005/0203142, published Sep. 15, 2005, which isincorporated herein by reference. Specific types of pain include, butare not limited to, nociceptive pain, neuropathic pain, mixed pain ofnociceptive and neuropathic pain, visceral pain, migraine, headache, andpost-operative pain.

Examples of nociceptive pain include, but are not limited to, painassociated with chemical or thermal burns, cuts of the skin, contusionsof the skin, osteoarthritis, rheumatoid arthritis, tendonitis, andmyofascial pain.

Examples of neuropathic pain include, but are not limited to, CRPS typeI. CRPS type II, reflex sympathetic dystrophy (RSD), reflexneurovascular dystrophy, reflex dystrophy, sympathetically maintainedpain syndrome, causalgia, Sudeck atrophy of bone, algoneurodystrophy,shoulder hand syndrome, post-traumatic dystrophy, trigeminal neuralgia,post herpetic neuralgia, cancer related pain, phantom limb pain,fibromyalgia, chronic fatigue syndrome, spinal cord injury pain, centralpost-stroke pain, radiculopathy, diabetic neuropathy, post-stroke pain,luetic neuropathy, and other painful neuropathic conditions such asthose induced by drugs such as vincristine and velcade. As used herein,the terms “complex regional pain syndrome,” “CRPS” and “CRPS and relatedsyndromes” mean a chronic pain disorder characterized by one or more ofthe following: pain, whether spontaneous or evoked, including allodynia(painful response to a stimulus that is not usually painful) andhyperalgesia (exaggerated response to a stimulus that is usually onlymildly painful); pain that is disproportionate to the inciting event(e.g., years of severe pain after an ankle sprain); regional pain thatis not limited to a single peripheral nerve distribution; and autonomicdysregulation (e.g., edema, alteration in blood flow and hyperhidrosis)associated with trophic skin changes (hair and nail growth abnormalitiesand cutaneous ulceration).

Examples of MD and related syndromes include, but are not limited to,those described in U.S. patent Publ. No. 2004/0091455, published May 13,2004, which is incorporated herein by reference. Specific examplesinclude, but are not limited to, atrophic (dry) MD, exudative (wet) MD,age-related maculopathy (ARM), choroidal neovascularisation (CNVM),retinal pigment epithelium detachment (PED), and atrophy of retinalpigment epithelium (RPE).

Examples of skin diseases include, but are not limited to, thosedescribed in U.S. Publ. No. 2005/0214328A1, published Sep. 29, 2005,which is incorporated herein by reference. Specific examples include,but are not limited to, keratoses and related symptoms, skin diseases ordisorders characterized with overgrowths of the epidermis, acne, andwrinkles. As used herein, the term “keratosis” refers to any lesion onthe epidermis marked by the presence of circumscribed overgrowths of thehorny layer, including but not limited to actinic keratosis, seborrheickeratosis, keratoacanthoma, keratosis follicularis (Darier disease),inverted follicular keratosis, palmoplantar keratoderma (PPK, keratosispalmaris et plantaris), keratosis pilaris, and stucco keratosis. Theterm “actinic keratosis” also refers to senile keratosis, keratosissenilis, verruca senilis, plana senilis, solar keratosis, keratoderma orkeratoma. The term “seborrheic keratosis” also refers to seborrheicwart, senile wart, or basal cell papilloma. Keratosis is characterizedby one or more of the following symptoms: rough appearing, scaly,erythematous papules, plaques, spicules or nodules on exposed surfaces(e.g., face, hands, ears, neck, legs and thorax), excrescences ofkeratin referred to as cutaneous horns, hyperkeratosis, telangiectasias,elastosis, pigmented lentigines, acanthosis, parakeratosis,dyskeratoses, papillomatosis, hyperpigmentation of the basal cells,cellular atypia, mitotic figures, abnormal cell-cell adhesion, denseinflammatory infiltrates and small prevalence of squamous cellcarcinomas.

Examples of skin diseases or disorders characterized with overgrowths ofthe epidermis include, but are not limited to, any conditions, diseasesor disorders marked by the presence of overgrowths of the epidermis,including but not limited to, infections associated with papillomavirus, arsenical keratoses, sign of Leser-Trélat, warty dyskeratoma(WD), trichostasis spinulosa (TS), erythrokeratodermia variabilis (EKV),ichthyosis fetalis (harlequin ichthyosis), knuckle pads, cutaneousmelanoacanthoma, porokeratosis, psoriasis, squamous cell carcinoma,confluent and reticulated papillomatosis (CRP), acrochordons, cutaneoushorn, cowden disease (multiple hamartoma syndrome), dermatosis papulosanigra (DPN), epidermal nevus syndrome (ENS), ichthyosis vulgaris,molluscum contagiosum, prurigo nodularis, and acanthosis nigricans (AN).

Examples of pulmonary disorders include, but are not limited to, thosedescribed in U.S. Publ. No. 2005/0239842A1, published Oct. 27, 2005,which is incorporated herein by reference. Specific examples includepulmonary hypertension and related disorders. Examples of pulmonaryhypertension and related disorders include, but are not limited to:primary pulmonary hypertension (PPH); secondary pulmonary hypertension(SPH); familial PPH; sporadic PPH; precapillary pulmonary hypertension;pulmonary arterial hypertension (PAH); pulmonary artery hypertension;idiopathic pulmonary hypertension; thrombotic pulmonary arteriopathy(TPA); plexogenic pulmonary arteriopathy; functional classes I to IVpulmonary hypertension; and pulmonary hypertension associated with,related to, or secondary to, left ventricular dysfunction, mitralvalvular disease, constrictive pericarditis, aortic stenosis,cardiomyopathy, mediastinal fibrosis, anomalous pulmonary venousdrainage, pulmonary venoocclusive disease, collagen vasular disease,congenital heart disease, HIV virus infection, drugs and toxins such asfenfluramines, congenital heart disease, pulmonary venous hypertension,chronic obstructive pulmonary disease, interstitial lung disease,sleep-disordered breathing, alveolar hypoventilation disorder, chronicexposure to high altitude, neonatal lung disease, alveolar-capillarydysplasia, sickle cell disease, other coagulation disorder, chronicthromboemboli, connective tissue disease, lupus including systemic andcutaneous lupus, schistosomiasis, sarcoidosis or pulmonary capillaryhemangiomatosis.

Examples of asbestos-related disorders include, but not limited to,those described in U.S. Publ. No. 2005/0100529, published May 12, 2005,which is incorporated herein by reference. Specific examples include,but are not limited to, mesothelioma, asbestosis, malignant pleuraleffusion, benign exudative effusion, pleural plaques, pleuralcalcification, diffuse pleural thickening, rounded atelectasis, fibroticmasses, and lung cancer.

Examples of parasitic diseases include, but are not limited to, thosedescribed in U.S. Publ. No. 2006/0154880, published Jul. 13, 2006, whichis incorporated herein by reference. Parasitic diseases include diseasesand disorders caused by human intracellular parasites such as, but notlimited to, P. falcifarium, P. ovale, P. vivax, P. malariae, L.donovari, L. infantum, L. aethiopica, L. major, L. tropica, L. mexicana,L. braziliensis, T. Gondii, B. microti, B. divergens, B. coli, C.parvum, C. cayetanensis, E. histolytica, I. belli, S. mansonii, S.haematobium, Trypanosoma ssp., Toxoplasma ssp., and O. volvulus. Otherdiseases and disorders caused by non-human intracellular parasites suchas, but not limited to, Babesia bovis, Babesia canis, Banesia Gibsoni,Besnoitia darlingi, Cytauxzoon felis, Eimeria ssp., Hammondia ssp., andTheileria ssp., are also encompassed. Specific examples include, but arenot limited to, malaria, babesiosis, trypanosomiasis, leishmaniasis,toxoplasmosis, meningoencephalitis, keratitis, amebiasis, giardiasis,cryptosporidiosis, isosporiasis, cyclosporiasis, microsporidiosis,ascariasis, trichuriasis, ancylostomiasis, strongyloidiasis,toxocariasis, trichinosis, lymphatic filariasis, onchocerciasis,filariasis, schistosomiasis, and dermatitis caused by animalschistosomes.

Examples of immunodeficiency disorders include, but are not limited to,those described in U.S. Publ. No. 2006/0188475, published Aug. 24, 2006,which is incorporated herein by reference. Specific examples include,but not limited to, adenosine deaminase deficiency, antibody deficiencywith normal or elevated Igs, ataxia-tenlangiectasia, bare lymphocytesyndrome, common variable immunodeficiency, Ig deficiency withhyper-IgM, Ig heavy chain deletions, IgA deficiency, immunodeficiencywith thymoma, reticular dysgenesis, Nezelof syndrome, selective IgGsubclass deficiency, transient hypogammaglobulinemia of infancy,Wistcott-Aldrich syndrome, X-linked agammaglobulinemia, X-linked severecombined immunodeficiency.

Examples of CNS disorders include, but are not limited to, thosedescribed in U.S. Publ. No. 2005/0143344A1, published Jun. 30, 2005,which is incorporated herein by reference. Specific examples include,but are not limited to, include, but are not limited to, AmyotrophicLateral Sclerosis, Alzheimer Disease, Parkinson Disease, Huntington'sDisease, Multiple Sclerosis other neuroimmunological disorders such asTourette Syndrome, delerium, or disturbances in consciousness that occurover a short period of time, and amnestic disorder, or discreet memoryimpairments that occur in the absence of other central nervous systemimpairments.

Examples of CNS injuries and related syndromes include, but are notlimited to, those described in U.S. Publ. No. 2006/0122228, publishedJun. 8, 2006, which is incorporated herein by reference. Specificexamples include, but are not limited to, CNS injury/damage and relatedsyndromes, include, but are not limited to, primary brain injury,secondary brain injury, traumatic brain injury, focal brain injury,diffuse axonal injury, head injury, concussion, post-concussionsyndrome, cerebral contusion and laceration, subdural hematoma,epidermal hematoma, post-traumatic epilepsy, chronic vegetative state,complete SCI, incomplete SCI, acute SCI, subacute SCI, chronic SCI,central cord syndrome, Brown-Sequard syndrome, anterior cord syndrome,conus medullaris syndrome, cauda equina syndrome, neurogenic shock,spinal shock, altered level of consciousness, headache, nausea, emesis,memory loss, dizziness, diplopia, blurred vision, emotional lability,sleep disturbances, irritability, inability to concentrate, nervousness,behavioral impairment, cognitive deficit, and seizure.

Other disease or disorders include, but not limited to, viral, genetic,allergic, and autoimmune diseases. Specific examples include, but notlimited to, HIV, hepatitis, adult respiratory distress syndrome, boneresorption diseases, chronic pulmonary inflammatory diseases,dermatitis, cystic fibrosis, septic shock, sepsis, endotoxic shock,hemodynamic shock, sepsis syndrome, post ischemic reperfusion injury,meningitis, psoriasis, fibrotic disease, cachexia, graft versus hostdisease, graft rejection, auto-immune disease, rheumatoid spondylitis,Crohn's disease, ulcerative colitis, inflammatory-bowel disease,multiple sclerosis, systemic lupus erythrematosus, ENL in leprosy,radiation damage, cancer, asthma, or hyperoxic alveolar injury.

Examples of atherosclerosis and related conditions include, but are notlimited to, those disclosed in U.S. Publ. No. 2002/0054899, publishedMay 9, 2002, which is incorporated herein by reference. Specificexamples include, but are not limited to, all forms of conditionsinvolving atherosclerosis, including restenosis after vascularintervention such as angioplasty, stenting, atherectomy and grafting.All forms of vascular intervention are contemplated by the disclosure,including diseases of the cardiovascular and renal system, such as, butnot limited to, renal angioplasty, percutaneous coronary intervention(PCI), percutaneous transluminal coronary angioplasty (PTCA), carotidpercutaneous transluminal angioplasty (PTA), coronary by-pass grafting,angioplasty with stent implantation, peripheral percutaneoustransluminal intervention of the iliac, femoral or popliteal arteries,and surgical intervention using impregnated artificial grafts. Thefollowing chart provides a listing of the major systemic arteries thatmay be in need of treatment, all of which are contemplated by thedisclosure:

Artery Body Area Supplied Axillary Shoulder and axilla Brachial Upperarm Brachiocephalic Head, neck, and arm Celiac Divides into leftgastric, splenic, and hepatic arteries Common carotid Neck Common iliacDivides into external and internal iliac arteries Coronary Heart Deepfemoral Thigh Digital Fingers Dorsalis pedis Foot External carotid Neckand external head regions External iliac Femoral artery Femoral ThighGastric Stomach Hepatic Liver, gallbladder, pancreas, and duodenumInferior mesenteric Descending colon, rectum, and pelvic wall Internalcarotid Neck and internal head regions Internal iliac Rectum, urinarybladder, external genitalia, buttocks muscles, uterus and vagina Leftgastric Esophagus and stomach Middle sacral Sacrum Ovarian OvariesPalmar arch Hand Peroneal Calf Popliteal Knee Posterior tibial CalfPulmonary Lungs Radial Forearm Renal Kidney Splenic Stomach, pancreas,and spleen Subclavian Shoulder Superior mesenteric Pancreas, smallintestine, ascending and transverse colon Testicular Testes UlnarForearm

Examples of dysfunctional sleep and related syndromes include, but arenot limited to, those disclosed in U.S. Publ. No. 2005/0222209A1,published Oct. 6, 2005, which is incorporated herein by reference.Specific examples include, but are not limited to, snoring, sleep apnea,insomnia, narcolepsy, restless leg syndrome, sleep terrors, sleepwalking sleep eating, and dysfunctional sleep associated with chronicneurological or inflammatory conditions. Chronic neurological orinflammatory conditions, include, but are not limited to, ComplexRegional Pain Syndrome, chronic low back pain, musculoskeletal pain,arthritis, radiculopathy, pain associated with cancer, fibromyalgia,chronic fatigue syndrome, visceral pain, bladder pain, chronicpancreatitis, neuropathies (diabetic, post-herpetic, traumatic orinflammatory), and neurodegenerative disorders such as Parkinson'sDisease, Alzheimer's Disease, amyotrophic lateral sclerosis, multiplesclerosis, Huntington's Disease, bradykinesia; muscle rigidity;parkinsonian tremor; parkinsonian gait; motion freezing; depression;defective long-term memory, Rubinstein-Taybi syndrome (RTS); dementia;postural instability; hypokinetic disorders; synuclein disorders;multiple system atrophies; striatonigral degeneration;olivopontocerebellar atrophy; Shy-Drager syndrome; motor neuron diseasewith parkinsonian features; Lewy body dementia; Tau pathology disorders;progressive supranuclear palsy; corticobasal degeneration;frontotemporal dementia; amyloid pathology disorders; mild cognitiveimpairment; Alzheimer disease with parkinsonism; Wilson disease;Hallervorden-Spatz disease; Chediak-Hagashi disease; SCA-3spinocerebellar ataxia; X-linked dystonia parkinsonism; prion disease;hyperkinetic disorders; chorea; ballismus; dystonia tremors; AmyotrophicLateral Sclerosis (ALS); CNS trauma and myoclonus.

Examples of hemoglobinopathy and related disorders include, but are notlimited to, those described in U.S. Publ. No. 2005/0143420A1, publishedJun. 30, 2005, which is incorporated herein by reference. Specificexamples include, but are not limited to, hemoglobinopathy, sickle cellanemia, and any other disorders related to the differentiation of CD34+cells.

Examples of TNFα and other cytokines related disorders include, but arenot limited to, those described in WO 98/03502 and WO 98/54170, both ofwhich are incorporated herein in their entireties by reference. Specificexamples include, but are not limited to: endotoxemia or toxic shocksyndrome; cachexia; adult respiratory distress syndrome; bone resorptiondiseases such as arthritis; hypercalcemia; Graft versus Host Reaction;cerebral malaria; inflammation; tumor growth; chronic pulmonaryinflammatory diseases: reperfusion injury; myocardial infarction;stroke; circulatory shock; rheumatoid arthritis; Crohn's disease; HIVinfection and AIDS; other disorders such as rheumatoid arthritis,rheumatoid spondylitis, osteoarthritis and other arthritic conditions,septic shock, septis, endotoxic shock, graft versus host disease,wasting, Crohn's disease, ulcerative colitis, multiple sclerosis,systemic lupus erythromatosis, ENL in leprosy, HIV, AIDS, andopportunistic infections in AIDS; cAMP related disorders such as septicshock, sepsis, endotoxic shock, hemodynamic shock and sepsis syndrome,post ischemic reperfusion injury, malaria, mycobacterial infection,meningitis, psoriasis, congestive heart failure, fibrotic disease,cachexia, graft rejection, oncogenic or cancerous conditions, asthma,autoimmune disease, radiation damages, and hyperoxic alveolar injury;viral infections, such as those caused by the herpes viruses; viralconjunctivitis; or atopic dermatitis.

In certain embodiments, the use of compounds provided herein in variousimmunological applications, in particular, as vaccine adjuvants,particularly anticancer vaccine adjuvants, as disclosed in U.S. Publ.No. 2007/0048327, published Mar. 1, 2007, which is incorporated hereinin its entirety by reference, is also encompassed. This aspect of thedisclosure also relates to the uses of compounds provided in combinationwith vaccines to treat or prevent cancer or infectious diseases, andother various uses of immunomodulatory compounds such as reduction ordesensitization of allergic reactions.

Depending on the condition, disorder, or disease to be treated and thesubject's condition, a compound provided herein may be administered byoral, parenteral (e.g., intramuscular, intraperitoneal, intravenous,ICV, intracistemal injection or infusion, subcutaneous injection, orimplant), inhalation, nasal, vaginal, rectal, sublingual, and/or topical(e.g., transdermal or local) routes of administration, and may beformulated alone or together in suitable dosage unit with apharmaceutically acceptable vehicle, carrier, diluent, excipient, or amixture thereof, appropriate for each route of administration.

The dose may be in the form of one, two, three, four, five, six, or moresub-doses that are administered at appropriate intervals per day. Thedose or sub-doses can be administered in the form of dosage unitscontaining from about 0.1 to about 1,000 mg, from about 0.1 to about 500mg, or from 0.5 about to about 100 mg of active ingredient(s) per dosageunit, and if the condition of the patient requires, the dose can, by wayof alternative, be administered as a continuous infusion.

In certain embodiments, an appropriate dosage level is about 0.01 toabout 100 mg per kg patient body weight per day (mg/kg per day), about0.01 to about 50 mg/kg per day, about 0.01 to about 25 mg/kg per day, orabout 0.05 to about 10 mg/kg per day, which may be administered insingle or multiple doses. A suitable dosage level may be about 0.01 toabout 100 mg/kg per day, about 0.05 to about 50 mg/kg per day, or about0.1 to about 10 mg/kg per day. Within this range, the dosage may beabout 0.01 to about 0.1, about 0.1 to about 1.0, about 1.0 to about 10,or about 10 to about 50 mg/kg per day.

For oral administration, the pharmaceutical compositions can be providedin the form of tablets containing 1.0 to 1,000 mg of the activeingredient, particularly about 1, about 5, about 10, about 15, about 20,about 25, about 50, about 75, about 100, about 150, about 200, about250, about 300, about 400, about 500, about 600, about 750, about 800,about 900, and about 1,000 mg of the active ingredient for thesymptomatic adjustment of the dosage to the patient to be treated. Thecompositions may be administered on a regimen of 1 to 4 times per day,including once, twice, three times, and four times per day.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, general health, sex, diet, mode and timeof administration, rate of excretion, drug combination, the severity ofthe particular condition, and the host undergoing therapy.

4.4 Second Active Agents

The compounds provided herein may also be combined or used incombination with other therapeutic agents useful in treating,preventing, and/or managing various diseases or disorders for which thecompounds provided herein are useful. It is believed that certaincombinations may work synergistically in the treatment of particulartypes of diseases or disorders, and conditions and symptoms associatedwith such diseases or disorders. A compound provided herein, including asingle enantiomer, a mixture of enantiomers, or a mixture ofdiastereomers thereof, or a pharmaceutically acceptable salt, solvate,or prodrug thereof, can also work to alleviate adverse effectsassociated with certain second active agents, and vice versa.

As used herein, the term “in combination” includes the use of more thanone therapeutic agents. However, the use of the term “in combination”does not restrict the order or route in which therapeutic agents areadministered to a subject with a condition, disorder, or disorder. Afirst therapeutic agent (e.g., a therapeutic agent such as a compoundprovided herein) can be administered prior to (e.g., 5 min, 15 min, 30min, 45 min, 1 hr, 2 hrs, 4 hrs, 6 hrs, 12 hrs, 24 hrs, 48 hrs, 72 hrs,96 hrs, 1 wk, 2 wks, 3 wks, 4 wks, 5 wks, 6 wks, 8 wks, or 12 wksbefore), concomitantly with, or subsequent to (e.g., 5 min, 15 min, 30min, 45 min, 1 hr, 2 hrs, 4 hrs, 6 hrs, 12 hrs, 24 hrs, 48 hrs, 72 hrs,96 hrs, 1 wk, 2 wks, 3 wks, 4 wks, 5 wks, 6 wks, 8 wks, or 12 wks after)the administration of a second therapeutic agent to a subject to betreated.

The suitability of a particular route of administration employed for aparticular active agent will depend on the active agent itself (e.g.,whether it can be administered orally without decomposing prior toentering the blood stream) and the disease being treated. In certainembodiments, the route of administration for compounds provided hereinis oral. In certain embodiments, the routes of administration for thesecond active agents or ingredients provided herein are those asdescribed in Physicians' Desk Reference, 1755-1760 (56th ed., 2002).

In one embodiment, the second active agent is administered intravenouslyor subcutaneously and once or twice daily in an amount of from about 1to about 1,000 mg, from about 5 to about 500 mg, from about 10 to about350 mg, or from about 50 to about 200 mg. The specific amount of thesecond active agent will depend on the specific agent used, the type ofdisease being treated or managed, the severity and stage of disease, andthe amount(s) of compounds provided herein and any optional additionalactive agents concurrently administered to the patient.

In certain embodiments, provided herein is a method of reducing,treating and/or preventing adverse or undesired effects associated withconventional therapy including, but not limited to, surgery,chemotherapy, radiation therapy, hormonal therapy, biological therapyand immunotherapy. Compounds provided herein and other activeingredients can be administered to a patient prior to, during, or afterthe occurrence of the adverse effect associated with conventionaltherapy.

When a compound provided herein is used contemporaneously with one ormore therapeutic agents, a pharmaceutical composition containing suchother agents in addition to the compound provided herein may beutilized, but is not required. Accordingly, the pharmaceuticalcompositions provided herein include those that also contain one or moreother therapeutic agents, in addition to a compound provided herein.

One or more second active ingredients or agents can be used in themethods and compositions provided herein. Second active agents can belarge molecules (e.g., proteins) or small molecules (e.g., syntheticinorganic, organometallic, or organic molecules).

Examples of large molecule active agents include, but are not limitedto, hematopoietic growth factors, cytokines, and monoclonal andpolyclonal antibodies. Specific examples of the active agents areanti-CD40 monoclonal antibodies (such as, for example, SGN-40,Herceptin, rituximab); histone deacetylyase inhibitors (such as, forexample, SAHA and LAQ 824); heat-shock protein-90 inhibitors (such as,for example, 17-AAG); insulin-like growth factor-I receptor kinaseinhibitors; vascular endothelial growth factor receptor kinaseinhibitors (such as, for example, PTK787); insulin growth factorreceptor inhibitors; lysophosphatidic acid acyltransrerase inhibitors;IkB kinase inhibitors; p38MAPK inhibitors; EGFR inhibitors (such as, forexample, gefitinib and erlotinib HCL); HER-2 antibodies (such as, forexample, trastuzumab (Herceptin®) and pertuzumab (Omnitarg™)); VEGFRantibodies (such as, for example, bevacizumab (Avastin™)); VEGFRinhibitors (such as, for example, flk-1 specific kinase inhibitors,SU5416 and ptk787/zk222584); PI3K inhibitors (such as, for example,wortmannin); C-Met inhibitors (such as, for example, PHA-665752);monoclonal antibodies (such as, for example, rituximab (Rituxan®),tositumomab (Bexxar®), edrecolomab (Panorex®) and G250); and anti-TNF-αantibodies. Examples of small molecule active agents include, but arenot limited to, small molecule anti-cancer agents and antibiotics (e.g.,clarithromycin).

Specific second active compounds that can be combined with compoundsprovided herein vary depending on the specific indication to be treated,prevented or managed.

For instance, for the treatment, prevention or management of cancer,second active agents include, but are not limited to: semaxanib;cyclosporin; etanercept; doxycycline; bortezomib; acivicin; aclarubicin;acodazole hydrochloride; acronine; adozelesin; aldesleukin; altretamine;ambomycin; ametantrone acetate; amsacrine; anastrozole; anthramycin;asparaginase; asperlin; azacitidine; azetepa; azotomycin; batimastat;benzodepa; bicalutamide; bisantrene hydrochloride; bisnafide dimesylate;bizelesin; bleomycin sulfate; brequinar sodium; bropirimine; busulfan;cactinomycin; calusterone; caracemide; carbetimer; carboplatin;carmustine; carubicin hydrochloride; carzelesin; cedefingol; celecoxib;chlorambucil; cirolemycin; cisplatin; cladribine; crisnatol mesylate;cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicinhydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguaninemesylate; diaziquone; docetaxel; doxorubicin; doxorubicin hydrochloride;droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin;edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin;enpromate; epipropidine; epirubicin hydrochloride; erbulozole;esorubicin hydrochloride; estramustine; estramustine phosphate sodium;etanidazole; etoposide; etoposide phosphate; etoprine; fadrozolehydrochloride; fazarabine; fenretinide; floxuridine; fludarabinephosphate; fluorouracil; fluorocitabine; fosquidone; fostriecin sodium;gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicinhydrochloride; ifosfamide; ilmofosine; iproplatin; irinotecan;irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolideacetate; liarozole hydrochloride; lometrexol sodium; lomustine;losoxantrone hydrochloride; masoprocol; maytansine; mechlorethaminehydrochloride; megestrol acetate; melengestrol acetate; melphalan;menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine;meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin;mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolicacid; nocodazole; nogalamycin; ormaplatin; oxisuran; paclitaxel;pegaspargase; peliomycin; pentamustine; peplomycin sulfate;perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine;procarbazine hydrochloride; puromycin; puromycin hydrochloride;pyrazofurin; riboprine; safingol; safingol hydrochloride; semustine;simtrazene; sparfosate sodium; sparsomycin; spirogermaniumhydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin;sulofenur; talisomycin; tecogalan sodium; taxotere; tegafur;teloxantrone hydrochloride; temoporfin; teniposide; teroxirone;testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;tirapazamine; toremifene citrate; trestolone acetate; triciribinephosphate; trimetrexate; trimetrexate glucuronate; triptorelin;tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;verteporfin; vinblastine sulfate; vincristine sulfate; vindesine;vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; and zorubicinhydrochloride.

Other second agents include, but are not limited to: 20-epi-1,25dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin;acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists;altretamine; ambamustine; amidox; amifostine; aminolevulinic acid;amrubiein; amsacrine; anagrelide; anastrozole; andrographolide;angiogenesis inhibitors; antagonist D; antagonist G; antarelix;anti-dorsalizing morphogenetic protein-1; antiandrogen, prostaticcarcinoma; antiestrogen; antineoplaston; antisense oligonucleotides;aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators;apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine;atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3;azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol;batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine;beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid;bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine;bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane;buthionine sulfoximine; calcipotriol; calphostin C; camptothecinderivatives; capecitabine; carboxamide-amino-triazole;carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor;carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropinB; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost;cis-porphyrin; cladribine; clomifene analogues; clotrimazole;collismycin A; collismycin B; combretastatin A4; combretastatinanalogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8;cryptophycin A derivatives; curacin A; cyclopentanthraquinones;cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor;cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin;dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone;didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;dihydrotaxol; dioxamycin; diphenyl spiromustine; docetaxel; docosanol;dolasetron; doxifluridine; doxorubicin; droloxifene; dronabinol;duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab;eflornithine; elemene; emitefur; epirubicin; epristeride; estramustineanalogue; estrogen agonists; estrogen antagonists; etanidazole;etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinide;filgrastim; finasteride; flavopiridol; flezelastine; fluasterone;fludarabine; fluorodaunorunicin hydrochloride; forfenimex; formestane;fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate;galocitabine; ganirelix; gelatinase inhibitors; gemcitabine; glutathioneinhibitors; hepsulfam; heregulin; hexamethylene bisacetamide; hypericin;ibandronic acid; idarubicin; idoxifene; idramantone; ilmofosine;ilomastat; imatinib (Gleevec), imiquimod; immunostimulant peptides;insulin-like growth factor-1 receptor inhibitor; interferon agonists;interferons; interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-;iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; loxoribine; lurtotecan; lutetiumtexaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A;marimastat; masoprocol; maspin; matrilysin inhibitors; matrixmetalloproteinase inhibitors; menogaril; merbarone; meterelin;methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine;mirimostim; mitoguazone; mitolactol; mitomycin analogues; mitonafide;mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene;molgramostim; Erbitux, human chorionic gonadotrophin; monophosphoryllipid A+myobacterium cell wall sk; mopidamol; mustard anticancer agent;mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; nilutamide; nisamycin; nitric oxidemodulators; nitroxide antioxidant; nitrullyn; oblimersen (Genasense®);O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid;panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase;peldesine; pentosan polysulfate sodium; pentostatin; pentrozole;perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rohitukine;romurtide; roquinimex; rubiginone B1; ruboxyl; safingol; saintopin;SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine;senescence derived inhibitor 1; sense oligonucleotides; signaltransduction inhibitors; sizofuran; sobuzoxane; sodium borocaptate;sodium phenylacetate; solverol; somatomedin binding protein; sonermin;sparfosic acid; spicamycin D; spiromustine; splenopentin; spongistatin1; squalamine; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; tallimustine; tamoxifen methiodide; tauromustine;tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomeraseinhibitors; temoporfin; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; translation inhibitors; tretinoin;triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron;turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors;ubenimex; urogenital sinus-derived growth inhibitory factor; urokinasereceptor antagonists; vapreotide; variolin B; velaresol; veramine;verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole;zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer.

Specific second active agents include, but are not limited to,2-methoxyestradiol, telomestatin, inducers of apoptosis in mutiplemyeloma cells (such as, for example, TRAIL), statins, semaxanib,cyclosporin, etanercept, doxycycline, bortezomib, oblimersen(Genasense®), remicade, docetaxel, celecoxib, melphalan, dexamethasone(Decadron®), steroids, gemcitabine, cisplatinum, temozolomide,etoposide, cyclophosphamide, temodar, carboplatin, procarbazine,gliadel, tamoxifen, topotecan, methotrexate, Arisa®, taxol, taxotere,fluorouracil, leucovorin, irinotecan, xeloda, CPT-11, interferon alpha,pegylated interferon alpha (e.g. PEG INTRON-A), capecitabine, cisplatin,thiotepa, fludarabine, carboplatin, liposomal daunorubicin, cytarabine,doxetaxol, pacilitaxel, vinblastine, IL-2, GM-CSF, dacarbazine,vinorelbine, zoledronic acid, palmitronate, biaxin, busulphan,prednisone, bisphosphonate, arsenic trioxide, vincristine, doxorubicin(Doxil®), paclitaxel, ganciclovir, adriamycin, estramustine sodiumphosphate (Emcyt®), sulindac, and etoposide.

Similarly, examples of specific second agents according to theindications to be treated, prevented, or managed can be found in thefollowing references, all of which are incorporated herein in theirentireties: U.S. Pat. Nos. 6,281,230 and 5,635,517; U.S. applicationSer. Nos. 10/411,649, 10/483,213, 10/411,656, 10/693,794, 10/699,154,and 10/981,189; and U.S. Prov. Appl. Nos. 60/554,923, 60/565,172,60/626,975, 60/630,599, 60/631,870, and 60/533,862.

Examples of second active agents that may be used for the treatment,prevention and/or management of pain include, but are not limited to,conventional therapeutics used to treat or prevent pain such asantidepressants, anticonvulsants, antihypertensives, anxiolytics,calcium channel blockers, muscle relaxants, non-narcotic analgesics,opioid analgesics, anti-inflammatories, cox-2 inhibitors,immunomodulatory agents, alpha-adrenergic receptor agonists orantagonists, immunosuppressive agents, corticosteroids, hyperbaricoxygen, ketamine, other anesthetic agents, NMDA antagonists, and othertherapeutics found, for example, in the Physician's Desk Reference 2003.Specific examples include, but are not limited to, salicylic acidacetate (Aspirin®), celecoxib (Celebrex®), Enbrel®, ketamine, gabapentin(Neurontin®), phenyloin (Dilantin®), carbamazepine (Tegretol®),oxcarbazepine (Trilepta®), valproic acid (Depakene®), morphine sulfate,hydromorphone, prednisone, griseofulvin, penthonium, alendronate,dyphenhydramide, guanethidine, ketorolac (Acular®), thyrocalcitonin,dimethylsulfoxide (DMSO), clonidine (Catapress®), bretylium, ketanserin,reserpine, droperidol, atropine, phentolamine, bupivacaine, lidocaine,acetaminophen, nortriptyline (Pamelor®), amitriptyline (Elavil®),imipramine (Tofranil®), doxepin (Sinequan®), clomipramine (Anafranil®),fluoxetine (Prozac), sertraline (Zoloft®), nefazodone (Serzone®),venlafaxine (Effexor®), trazodone (Desyrel®), bupropion (Wellbutrin®),mexiletine, nifedipine, propranolol, tramadol, lamotrigine, ziconotide,ketamine, dextromethorphan, benzodiazepines, baclofen, tizanidine andphenoxybenzamine.

Examples of second active agents that may be used for the treatment,prevention and/or management of MD and related syndromes include, butare not limited to, a steroid, a light sensitizer, an integrin, anantioxidant, an interferon, a xanthine derivative, a growth hormone, aneutrotrophic factor, a regulator of neovascularization, an anti-VEGFantibody, a prostaglandin, an antibiotic, a phytoestrogen, ananti-inflammatory compound or an antiangiogenesis compound, or acombination thereof. Specific examples include, but are not limited to,verteporfin, purlytin, an angiostatic steroid, rhuFab, interferon-2a,pentoxifylline, tin etiopurpurin, motexafin lutetium,9-fluoro-11,21-dihydroxy-16,17-1-methylethylidine-bis(oxy)pregna-1,4-diene-3,20-dione, latanoprost(see U.S. Pat. No. 6,225,348), tetracycline and its derivatives,rifamycin and its derivatives, macrolides, metronidazole (U.S. Pat. Nos.6,218,369 and 6,015,803), genistein, genistin, 6′-O-Mal genistin,6′-O-Ac genistin, daidzein, daidzin, 6′-O-Mal daidzin, 6′-O-Ac daidzin,glycitein, glycitin, 6′-O-Mal glycitin, biochanin A, formononetin (U.S.Pat. No. 6,001,368), triamcinolone acetomide, dexamethasone (U.S. Pat.No. 5,770,589), thalidomide, glutathione (U.S. Pat. No. 5,632,984),basic fibroblast growth factor (bFGF), transforming growth factor b(TGF-b), brain-derived neurotrophic factor (BDNF), plasminogen activatorfactor type 2 (PAI-2), EYE101 (Eyetech Pharmaceuticals), LY333531 (EliLilly), Miravant, and RETISERT implant (Bausch & Lomb). All of thereferences cited above are incorporated herein in their entireties byreference.

Examples of second active agents that may be used for the treatment,prevention and/or management of skin diseases include, but are notlimited to, keratolytics, retinoids, α-hydroxy acids, antibiotics,collagen, botulinum toxin, interferon, and immunomodulatory agents.Specific examples include, but are not limited to, 5-fluorouracil,masoprocol, trichloroacetic acid, salicylic acid, lactic acid, ammoniumlactate, urea, tretinoin, isotretinoin, antibiotics, collagen, botulinumtoxin, interferon, corticosteroid, transretinoic acid and collagens suchas human placental collagen, animal placental collagen, Dermalogen,AlloDerm, Fascia, Cymetra, Autologen, Zyderm, Zyplast, Resoplast, andIsolagen.

Examples of second active agents that may be used for the treatment,prevention and/or management of pulmonary hepertension and relateddisorders include, but are not limited to, anticoagulants, diuretics,cardiac glycosides, calcium channel blockers, vasodilators, prostacyclinanalogues, endothelin antagonists, phosphodiesterase inhibitors (e.g.,PDE V inhibitors), endopeptidase inhibitors, lipid lowering agents,thromboxane inhibitors, and other therapeutics known to reduce pulmonaryartery pressure. Specific examples include, but are not limited to,warfarin (Coumadin®), a diuretic, a cardiac glycoside, digoxin-oxygen,diltiazem, nifedipine, a vasodilator such as prostacyclin (e.g.,prostaglandin I2 (PGI2), epoprostenol (EPO, Floran®), treprostinil(Remodulin®), nitric oxide (NO), bosentan (Tracleer®), amlodipine,epoprostenol (Floran®), treprostinil (Remodulin®), prostacyclin,tadalafil (Clalis®), simvastatin (Zocor®), omapatrilat (Vanlev®),irbesartan (Avapro®), pravastatin (Pravachol®), digoxin, L-arginine,iloprost, betaprost, and sildenafil (Viagra®).

Examples of second active agents that may be used for the treatment,prevention and/or management of asbestos-related disorders include, butare not limited to, anthracycline, platinum, alkylating agent,oblimersen (Genasense®), cisplatinum, cyclophosphamide, temodar,carboplatin, procarbazine, gliadel, tamoxifen, topotecan, methotrexate,taxotere, irinotecan, capecitabine, cisplatin, thiotepa, fludarabine,carboplatin, liposomal daunorubicin, cytarabine, doxetaxol, pacilitaxel,vinblastine, IL-2, GM-CSF, dacarbazine, vinorelbine, zoledronic acid,palmitronate, biaxin, busulphan, prednisone, bisphosphonate, arsenictrioxide, vincristine, doxorubicin (Doxil®), paclitaxel, ganciclovir,adriamycin, bleomycin, hyaluronidase, mitomycin C, mepacrine, thiotepa,tetracycline and gemcitabine.

Examples of second active agents that may be used for the treatment,prevention and/or management of parasitic diseases include, but are notlimited to, chloroquine, quinine, quinidine, pyrimethamine,sulfadiazine, doxycycline, clindamycin, mefloquine, halofantrine,primaquine, hydroxychloroquine, proguanil, atovaquone, azithromycin,suramin, pentamidine, melarsoprol, nifurtimox, benznidazole,amphotericin B, pentavalent antimony compounds (e.g., sodiumstiboglucuronate), interfereon gamma, itraconazole, a combination ofdead promastigotes and BCG, leucovorin, corticosteroids, sulfonamide,spiramycin, IgG (serology), trimethoprim, and sulfamethoxazole.

Examples of second active agents that may be used for the treatment,prevention and/or management of immunodeficiency disorders include, butare not limited to: antibiotics (therapeutic or prophylactic) such as,but not limited to, ampicillin, clarithromycin, tetracycline,penicillin, cephalosporins, streptomycin, kanamycin, and erythromycin;antivirals such as, but not limited to, amantadine, rimantadine,acyclovir, and ribavirin; immunoglobulin; plasma; immunologic enhancingdrugs such as, but not limited to, levamisole and isoprinosine;biologics such as, but not limited to, gammaglobulin, transfer factor,interleukins, and interferons; hormones such as, but not limited to,thymic; and other immunologic agents such as, but not limited to, B cellstimulators (e.g., BAFF/BlyS), cytokines (e.g., IL-2, IL-4, and IL-5),growth factors (e.g., TGF-α), antibodies (e.g., anti-CD40 and IgM),oligonucleotides containing unmethylated CpG motifs, and vaccines (e.g.,viral and tumor peptide vaccines).

Examples of second active agents that may be used for the treatment,prevention and/or management of CNS disorders include, but are notlimited to: a dopamine agonist or antagonist, such as, but not limitedto, Levodopa, L-DOPA, cocaine, α-methyl-tyrosine, reserpine,tetrabenazine, benzotropine, pargyline, fenodolpam mesylate,cabergoline, pramipexole dihydrochloride, ropinorole, amantadinehydrochloride, selegiline hydrochloride, carbidopa, pergolide mesylate,Sinemet CR, and Symmetrel; a MAO inhibitor, such as, but not limited to,iproniazid, clorgyline, phenelzine and isocarboxazid; a COMT inhibitor,such as, but not limited to, tolcapone and entacapone; a cholinesteraseinhibitor, such as, but not limited to, physostigmine saliclate,physostigmine sulfate, physostigmine bromide, meostigmine bromide,neostigmine methylsulfate, ambenonim chloride, edrophonium chloride,tacrine, pralidoxime chloride, obidoxime chloride, trimedoxime bromide,diacetyl monoxim, endrophonium, pyridostigmine, and demecarium; ananti-inflammatory agent, such as, but not limited to, naproxen sodium,diclofenac sodium, diclofenac potassium, celecoxib, sulindac, oxaprozin,diflunisal, etodolac, meloxicam, ibuprofen, ketoprofen, nabumetone,refecoxib, methotrexate, leflunomide, sulfasalazine, gold salts, Rho-DImmune Globulin, mycophenylate mofetil, cyclosporine, azathioprine,tacrolimus, basiliximab, daclizumab, salicylic acid, acetylsalicylicacid, methyl salicylate, diflunisal, salsalate, olsalazine,sulfasalazine, acetaminophen, indomethacin, sulindac, mefenamic acid,meclofenamate sodium, tolmetin, ketorolac, dichlofenac, flurbinprofen,oxaprozin, piroxicam, meloxicam, ampiroxicam, droxicam, pivoxicam,tenoxicam, phenylbutazone, oxyphenbutazone, antipyrine, aminopyrine,apazone, zileuton, aurothioglucose, gold sodium thiomalate, auranofin,methotrexate, colchicine, allopurinol, probenecid, sulfinpyrazone andbenzbromarone or betamethasone and other glucocorticoids; and anantiemetic agent, such as, but not limited to, metoclopromide,domperidone, prochlorperazine, promethazine, chlorpromazine,trimethobenzamide, ondansetron, granisetron, hydroxyzine, acetylleucinemonoethanolamine, alizapride, azasetron, benzquinamide, bietanautine,bromopride, buclizine, clebopride, cyclizine, dimenhydrinate,diphenidol, dolasetron, meclizine, methallatal, metopimazine, nabilone,oxyperndyl, pipamazine, scopolamine, sulpiride, tetrahydrocannabinol,thiethylperazine, thioproperazine, tropisetron, and a mixture thereof.

Examples of second active agents that may be used for the treatment,prevention and/or management of CNS injuries and related syndromesinclude, but are not limited to, immunomodulatory agents,immunosuppressive agents, antihypertensives, anticonvulsants,fibrinolytic agents, antiplatelet agents, antipsychotics,antidepressants, benzodiazepines, buspirone, amantadine, and other knownor conventional agents used in patients with CNS injury/damage andrelated syndromes. Specific examples include, but are not limited to:steroids (e.g., glucocorticoids, such as, but not limited to,methylprednisolone, dexamethasone and betamethasone); ananti-inflammatory agent, including, but not limited to, naproxen sodium,diclofenac sodium, diclofenac potassium, celecoxib, sulindac, oxaprozin,diflunisal, etodolac, meloxicam, ibuprofen, ketoprofen, nabumetone,refecoxib, methotrexate, leflunomide, sulfasalazine, gold salts, RHo-DImmune Globulin, mycophenylate mofetil, cyclosporine, azathioprine,tacrolimus, basiliximab, daclizumab, salicylic acid, acetylsalicylicacid, methyl salicylate, diflunisal, salsalate, olsalazine,sulfasalazine, acetaminophen, indomethacin, sulindac, mefenamic acid,meclofenamate sodium, tolmetin, ketorolac, dichlofenac, flurbinprofen,oxaprozin, piroxicam, meloxicam, ampiroxicam, droxicam, pivoxicam,tenoxicam, phenylbutazone, oxyphenbutazone, antipyrine, aminopyrine,apazone, zileuton, aurothioglucose, gold sodium thiomalate, auranofin,methotrexate, colchicine, allopurinol, probenecid, sulfinpyrazone andbenzbromarone; a cAMP analog including, but not limited to, db-cAMP; anagent comprising a methylphenidate drug, which comprises1-threo-methylphenidate, d-threo-methylphenidate,dl-threo-methylphenidate, 1-erythro-methylphenidate,d-erythro-methylphenidate, dl-erythro-methylphenidate, and a mixturethereof; and a diuretic agent such as, but not limited to, mannitol,furosemide, glycerol, and urea.

Examples of second active agent that may be used for the treatment,prevention and/or management of dysfunctional sleep and relatedsyndromes include, but are not limited to, a tricyclic antidepressantagent, a selective serotonin reuptake inhibitor, an antiepileptic agent(gabapentin, pregabalin, carbamazepine, oxcarbazepine, levitiracetam,topiramate), an antiaryhthmic agent, a sodium channel blocking agent, aselective inflammatory mediator inhibitor, an opioid agent, a secondimmunomodulatory compound, a combination agent, and other known orconventional agents used in sleep therapy. Specific examples include,but are not limited to, Neurontin, oxycontin, morphine, topiramate,amitryptiline, nortryptiline, carbamazepine, Levodopa, L-DOPA, cocaine,α-methyl-tyrosine, reserpine, tetrabenazine, benzotropine, pargyline,fenodolpam mesylate, cabergoline, pramipexole dihydrochloride,ropinorole, amantadine hydrochloride, selegiline hydrochloride,carbidopa, pergolide mesylate, Sinemet CR, Symmetrel, iproniazid,clorgyline, phenelzine, isocarboxazid, tolcapone, entacapone,physostigmine saliclate, physostigmine sulfate, physostigmine bromide,meostigmine bromide, neostigmine methylsulfate, ambenonim chloride,edrophonium chloride, tacrine, pralidoxime chloride, obidoxime chloride,trimedoxime bromide, diacetyl monoxim, endrophonium, pyridostigmine,demecarium, naproxen sodium, diclofenac sodium, diclofenac potassium,celecoxib, sulindac, oxaprozin, diflunisal, etodolac, meloxicam,ibuprofen, ketoprofen, nabumetone, refecoxib, methotrexate, leflunomide,sulfasalazine, gold salts, RHo-D Immune Globulin, mycophenylate mofetil,cyclosporine, azathioprine, tacrolimus, basiliximab, daclizumab,salicylic acid, acetylsalicylic acid, methyl salicylate, diflunisal,salsalate, olsalazine, sulfasalazine, acetaminophen, indomethacin,sulindac, mefenamic acid, meclofenamate sodium, tolmetin, ketorolac,dichlofenac, flurbinprofen, oxaprozin, piroxicam, meloxicam,ampiroxicam, droxicam, pivoxicam, tenoxicam, phenylbutazone,oxyphenbutazone, antipyrine, aminopyrine, apazone, zileuton,aurothioglucose, gold sodium thiomalate, auranofin, methotrexate,colchicine, allopurinol, probenecid, sulfinpyrazone, benzbromarone,betamethasone and other glucocorticoids, metoclopromide, domperidone,prochlorperazine, promethazine, chlorpromazine, trimethobenzamide,ondansetron, granisetron, hydroxyzine, acetylleucine monoethanolamine,alizapride, azasetron, benzquinamide, bietanautine, bromopride,buclizine, clebopride, cyclizine, dimenhydrinate, diphenidol,dolasetron, meclizine, methallatal, metopimazine, nabilone, oxyperndyl,pipamazine, scopolamine, sulpiride, tetrahydrocannabinol,thiethylperazine, thioproperazine, tropisetron, and a mixture thereof.

Examples of second active agents that may be used for the treatment,prevention and/or management of hemoglobinopathy and related disordersinclude, but are not limited to: interleukins, such as IL-2 (includingrecombinant IL-II (“rIL2”) and canarypox IL-2), IL-10, IL-12, and IL-18;interferons, such as interferon alfa-2a, interferon alfa-2b, interferonalfa-n1, interferon alfa-n3, interferon beta-I a, and interferon gamma-Ib; and G-CSF; hydroxyurea; butyrates or butyrate derivatives: nitrousoxide; HEMOXIN™ (NIPRISAN™; see U.S. Pat. No. 5,800,819); Gardos channelantagonists such as clotrimazole and triaryl methane derivatives;Deferoxamine; protein C; and transfusions of blood, or of a bloodsubstitute such as Hemospan™ or Hemospan™ PS (Sangart).

The dosages given will depend on absorption, inactivation and excretionrates of the drug as well as other factors known to those of skill inthe art. It is to be noted that dosage values will also vary with theseverity of the condition to be alleviated. It is to be furtherunderstood that for any particular subject, specific dosage regimens andschedules should be adjusted over time according to the individual needand the professional judgment of the person administering or supervisingthe administration of the compositions.

The weight ratio of a compound provided herein to the second activeingredient depends upon the effective dose of each ingredient.Generally, an effective dose of each will be used. Thus, for example,when a compound provided herein is combined with a PPAR agonist theweight ratio of the compound provided herein to the PPAR agonist willgenerally range from about 1000:1 to about 1:1000 or about 200:1 toabout 1:200. Combinations of a compound provided herein and other activeingredients will generally also be within the aforementioned range, butin each case, an effective dose of each active ingredient should beused.

4.5 Cycling Therapy

In certain embodiments, the prophylactic or therapeutic agents providedherein are cyclically administered to a patient. Cycling therapyinvolves the administration of an active agent for a period of time,followed by a rest for a period of time, and repeating this sequentialadministration. Cycling therapy can reduce the development of resistanceto one or more of the therapies, avoid or reduce the side effects of oneof the therapies, and/or improves the efficacy of the treatment.

In certain embodiments, a compound provided herein is administered dailyin a single or divided dose in a four to six week cycle with a restperiod of about a week or two weeks. The disclosure further allows thefrequency, number, and length of dosing cycles to be increased. Incertain embodiments, a compound provided herein is administered for morecycles than are typical when it is administered alone. In certainembodiments, a compound provided herein is administered for a greaternumber of cycles that would typically cause dose-limiting toxicity in apatient to whom a second active ingredient is not also beingadministered.

In one embodiment, a compound provided herein is administered daily andcontinuously for three or four weeks at a dose of from about 0.1 mg toabout 500 mg per day, followed by a break of one or two weeks. In otherembodiments, the dose can be from about 1 mg to about 300 mg, from about0.1 mg to about 150 mg, from about 1 mg to about 200 mg, from about 10mg to about 100 mg, from about 0.1 mg to about 50 mg, from about 1 mg toabout 50 mg, from about 10 mg to about 50 mg, from about 20 mg to about30 mg, or from about 1 mg to about 20 mg, followed by a break.

In one embodiment, a compound provided herein and a second activeingredient are administered orally, with administration of the compoundprovided herein occurring 30 to 60 minutes prior to the second activeingredient, during a cycle of four to six weeks. In another embodiment,the combination of a compound provided herein and a second activeingredient is administered by intravenous infusion over about 90 minutesevery cycle. In certain embodiments, the number of cycles during whichthe combinatorial treatment is administered to a patient is from aboutone to about 24 cycles, from about two to about 16 cycles, or from aboutfour to about three cycles.

4.6 Pharmaceutical Compositions and Dosage Forms

Pharmaceutical compositions can be used in the preparation ofindividual, single unit dosage forms. Pharmaceutical compositions anddosage forms provided herein comprise a compound provided herein, or apharmaceutically acceptable salt, solvate, stereoisomer, or prodrugthereof. Pharmaceutical compositions and dosage forms provided hereincan further comprise one or more excipients.

Pharmaceutical compositions and dosage forms provided herein can alsocomprise one or more additional active ingredients. Examples of optionalsecond, or additional, active ingredients are disclosed in Section 4.4,above.

Single unit dosage forms provided herein are suitable for oral, mucosal(e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (e.g.,subcutaneous, intravenous, bolus injection, intramuscular, orintraarterial), topical (e.g., eye drops or other ophthalmicpreparations), transdermal or transcutaneous administration to apatient. Examples of dosage forms include, but are not limited to:tablets; caplets; capsules, such as soft elastic gelatin capsules;cachets; troches; lozenges; dispersions; suppositories; powders;aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage formssuitable for oral or mucosal administration to a patient, includingsuspensions (e.g., aqueous or non-aqueous liquid suspensions,oil-in-water emulsions, or a water-in-oil liquid emulsions), solutions,and elixirs; liquid dosage forms suitable for parenteral administrationto a patient; eye drops or other ophthalmic preparations suitable fortopical administration; and sterile solids (e.g., crystalline oramorphous solids) that can be reconstituted to provide liquid dosageforms suitable for parenteral administration to a patient.

The composition, shape, and type of dosage forms provided herein willtypically vary depending on their use. For example, a dosage form usedin the acute treatment of a disease may contain larger amounts of one ormore of the active ingredients it comprises than a dosage form used inthe chronic treatment of the same disease. Similarly, a parenteraldosage form may contain smaller amounts of one or more of the activeingredients it comprises than an oral dosage form used to treat the samedisease. These and other ways in which specific dosage forms encompassedby this disclosure will vary from one another will be readily apparentto those skilled in the art. See, e.g., Remington's PharmaceuticalSciences, 18th ed., Mack Publishing, Easton Pa. (1990).

In certain embodiments, the pharmaceutical compositions and dosage formsprovided herein comprise one or more excipients. Suitable excipients arewell known to those skilled in the art of pharmacy, and non-limitingexamples of suitable excipients are provided herein. Whether aparticular excipient is suitable for incorporation into a pharmaceuticalcomposition or dosage form depends on a variety of factors well known inthe art including, but not limited to, the way in which the dosage formwill be administered to a patient. For example, oral dosage forms suchas tablets may contain excipients not suited for use in parenteraldosage forms. The suitability of a particular excipient may also dependon the specific active ingredients in the dosage form. For example, thedecomposition of some active ingredients may be accelerated by someexcipients such as lactose, or when exposed to water. Active ingredientsthat comprise primary or secondary amines are particularly susceptibleto such accelerated decomposition. Consequently, this disclosureencompasses pharmaceutical compositions and dosage forms that containlittle, if any, lactose other mono- or di-saccharides. As used herein,the term “lactose-free” means that the amount of lactose present, ifany, is insufficient to substantially increase the degradation rate ofan active ingredient.

Lactose-free compositions provided herein can comprise excipients thatare well known in the art and are listed, for example, in the U.S.Pharmacopeia (USP) 25-NF20 (2002). In general, lactose-free compositionscomprise active ingredients, a binder/filler, and a lubricant inpharmaceutically compatible and pharmaceutically acceptable amounts. Incertain embodiments, lactose-free dosage forms provided herein compriseactive ingredients, microcrystalline cellulose, pre-gelatinized starch,and magnesium stearate.

This disclosure further encompasses anhydrous pharmaceuticalcompositions and dosage forms comprising active ingredients, since watercan facilitate the degradation of some compounds. For example, theaddition of water (e.g., 5%) is widely accepted in the pharmaceuticalarts as a means of simulating long-term storage in order to determinecharacteristics such as shelf-life or the stability of formulations overtime. See, e.g., Jens T. Carstensen, Drug Stability: Principles &Practice. 2d. Ed., Marcel Dekker, NY, N.Y., 1995, pp. 379-80. In effect,water and heat accelerate the decomposition of some compounds. Thus, theeffect of water on a formulation can be of great significance sincemoisture and/or humidity are commonly encountered during manufacture,handling, packaging, storage, shipment, and use of formulations.

Anhydrous pharmaceutical compositions and dosage forms provided hereincan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions. In certain embodiments,pharmaceutical compositions and dosage forms that comprise lactose andat least one active ingredient that comprises a primary or secondaryamine are anhydrous if substantial contact with moisture and/or humidityduring manufacturing, packaging, and/or storage is expected.

An anhydrous pharmaceutical composition should be prepared and storedsuch that its anhydrous nature is maintained. In certain embodiments,anhydrous compositions are packaged using materials known to preventexposure to water such that they can be included in suitable formularykits. Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastics, unit dose containers (e.g., vials),blister packs, and strip packs.

The disclosure further encompasses pharmaceutical compositions anddosage forms that comprise one or more compounds that reduce the rate bywhich an active ingredient will decompose. Such compounds, which arereferred to herein as “stabilizers,” include, but are not limited to,antioxidants such as ascorbic acid, pH buffers, or salt buffers. Likethe amounts and types of excipients, the amounts and specific types ofactive ingredients in a dosage form may differ depending on factors suchas, but not limited to, the route by which it is to be administered topatients. In certain embodiments, dosage forms provided herein comprisea compound provided herein in an amount of from about 0.10 to about 500mg. In certain embodiments, the dosage forms provided herein comprise acompound provided herein in an amount of about 0.1, 1, 2, 5, 7.5, 10,12.5, 15, 17.5, 20, 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, or500 mg.

In certain embodiments, the dosage forms comprise the second activeingredient in an amount of 1 to about 1,000 mg, from about 5 to about500 mg, from about 10 to about 350 mg, or from about 50 to about 200 mg.Of course, the specific amount of the second active agent will depend onthe specific agent used, the type of diseases being treated or managed,and the amount(s) of a compound provided herein and any optionaladditional active agents concurrently administered to the patient.

4.6.1 Oral Dosage Forms

Pharmaceutical compositions provided herein that are suitable for oraladministration can be presented as discrete dosage forms, such as, butare not limited to, tablets (e.g., chewable tablets), caplets, capsules,and liquids (e.g., flavored syrups). Such dosage forms containpredetermined amounts of active ingredients, and may be prepared bymethods of pharmacy well known to those skilled in the art. Seegenerally, Remington's Pharmaceutical Sciences, 18th ed., MackPublishing, Easton Pa. (1990).

In certain embodiments, oral dosage forms provided herein are preparedby combining the active ingredients in an intimate admixture with atleast one excipient according to conventional pharmaceutical compoundingtechniques. Excipients can take a wide variety of forms depending on theform of preparation desired for administration. For example, excipientssuitable for use in oral liquid or aerosol dosage forms include, but arenot limited to, water, glycols, oils, alcohols, flavoring agents,preservatives, and coloring agents. Examples of excipients suitable foruse in solid oral dosage forms (e.g., powders, tablets, capsules, andcaplets) include, but are not limited to, starches, sugars,micro-crystalline cellulose, diluents, granulating agents, lubricants,binders, and disintegrating agents. Because of their ease ofadministration, tablets and capsules represent the most advantageousoral dosage unit forms, in which case solid excipients are employed. Ifdesired, tablets can be coated by standard aqueous or nonaqueoustechniques. Such dosage forms can be prepared by any of the methods ofpharmacy. In general, pharmaceutical compositions and dosage forms areprepared by uniformly and intimately admixing the active ingredientswith liquid carriers, finely divided solid carriers, or both, and thenshaping the product into the desired presentation if necessary.

For example, a tablet can be prepared by compression or molding.Compressed tablets can be prepared by compressing in a suitable machinethe active ingredients in a free-flowing form such as powder orgranules, optionally mixed with an excipient. Molded tablets can be madeby molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

Examples of excipients that can be used in oral dosage forms providedherein include, but are not limited to, binders, tillers, disintegrants,and lubricants. Binders suitable for use in pharmaceutical compositionsand dosage forms include, but are not limited to, corn starch, potatostarch, or other starches, gelatin, natural and synthetic gums such asacacia, sodium alginate, alginic acid, other alginates, powderedtragacanth, guar gum, cellulose and its derivatives (e.g., ethylcellulose, cellulose acetate, carboxymethyl cellulose calcium, sodiumcarboxymethyl cellulose), polyvinyl pyrrolidone, methyl cellulose,pre-gelatinized starch, hydroxypropyl methyl cellulose, (e.g., Nos.2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof.

Suitable forms of microcrystalline cellulose include, but are notlimited to, the materials sold as AVICEL-PH-101, AVICEL-PH-103 AVICELRC-581, AVICEL-PH-105 (available from FMC Corporation, American ViscoseDivision, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof. Anspecific binder is a mixture of microcrystalline cellulose and sodiumcarboxymethyl cellulose sold as AVICEL RC-581. Suitable anhydrous or lowmoisture excipients or additives include AVICEL-PH-103™ and Starch 1500LM.

Examples of fillers suitable for use in the pharmaceutical compositionsand dosage forms provided herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof. Incertain embodiments, the binder or filler in pharmaceutical compositionsprovided herein is present in from about 50 to about 99 weight percentof the pharmaceutical composition or dosage form.

Disintegrants are used in the compositions provided herein to providetablets that disintegrate when exposed to an aqueous environment.Tablets that contain too much disintegrant may disintegrate in storage,while those that contain too little may not disintegrate at a desiredrate or under the desired conditions. Thus, a sufficient amount ofdisintegrant that is neither too much nor too little to detrimentallyalter the release of the active ingredients should be used to form solidoral dosage forms provided herein. The amount of disintegrant usedvaries based upon the type of formulation, and is readily discernible tothose of ordinary skill in the art. In certain embodiments,pharmaceutical compositions provided herein comprise from about 0.5 toabout 15 weight percent of disintegrant, or from about 1 to about 5weight percent of disintegrant.

Disintegrants that can be used in pharmaceutical compositions and dosageforms provided herein include, but are not limited to, agar-agar,alginic acid, calcium carbonate, microcrystalline cellulose,croscarmellose sodium, crospovidone, polacrilin potassium, sodium starchglycolate, potato or tapioca starch, other starches, pre-gelatinizedstarch, other starches, clays, other algins, other celluloses, gums, andmixtures thereof.

Lubricants that can be used in pharmaceutical compositions and dosageforms provided herein include, but are not limited to, calcium stearate,magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol,mannitol, polyethylene glycol, other glycols, stearic acid, sodiumlauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut oil,cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, andmixtures thereof. Additional lubricants include, for example, a syloidsilica gel (AEROSIL200, manufactured by W.R. Grace Co. of Baltimore,Md.), a coagulated aerosol of synthetic silica (marketed by Degussa Co.of Plano, Tex.), CAB-O-SIL (a pyrogenic silicon dioxide product sold byCabot Co. of Boston, Mass.), and mixtures thereof. If used at all,lubricants are typically used in an amount of less than about 1 weightpercent of the pharmaceutical compositions or dosage forms into whichthey are incorporated.

A solid oral dosage form provided herein comprises a compound providedherein, anhydrous lactose, microcrystalline cellulose,polyvinylpyrrolidone, stearic acid, colloidal anhydrous silica, andgelatin.

4.6.2 Controlled Release Dosage Forms

Active ingredients provided herein can be administered by controlledrelease means or by delivery devices that are well known to those ofordinary skill in the art. Examples include, but are not limited to,those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809;3,598,123; and 4,008,719, 5,674,533, 5,059,595, 5,591,767, 5,120,548,5,073,543, 5,639,476, 5,354,556, and 5,733,566, each of which isincorporated herein by reference. Such dosage forms can be used toprovide slow or controlled-release of one or more active ingredientsusing, for example, hydropropylmethyl cellulose, other polymer matrices,gels, permeable membranes, osmotic systems, multilayer coatings,microparticles, liposomes, microspheres, or a combination thereof toprovide the desired release profile in varying proportions. Suitablecontrolled-release formulations known to those of ordinary skill in theart, including those described herein, can be readily selected for usewith the active ingredients provided herein. The disclosure thusencompasses single unit dosage forms suitable for oral administrationsuch as, but not limited to, tablets, capsules, gelcaps, and capletsthat are adapted for controlled-release.

All controlled-release pharmaceutical products have a common goal ofimproving drug therapy over that achieved by their non-controlledcounterparts. Ideally, the use of an optimally designedcontrolled-release preparation in medical treatment is characterized bya minimum of drug substance being employed to cure or control thecondition in a minimum amount of time. Advantages of controlled-releaseformulations include extended activity of the drug, reduced dosagefrequency, and increased patient compliance. In addition,controlled-release formulations can be used to affect the time of onsetof action or other characteristics, such as blood levels of the drug,and can thus affect the occurrence of side (e.g., adverse) effects.

Most controlled-release formulations are designed to initially releasean amount of drug (active ingredient) that promptly produces the desiredtherapeutic effect, and gradually and continually release of otheramounts of drug to maintain this level of therapeutic or prophylacticeffect over an extended period of time. In order to maintain thisconstant level of drug in the body, the drug must be released from thedosage form at a rate that will replace the amount of drug beingmetabolized and excreted from the body. Controlled-release of an activeingredient can be stimulated by various conditions including, but notlimited to, pH, temperature, enzymes, water, or other physiologicalconditions or compounds.

4.6.3 Parenteral Dosage Forms

Parenteral dosage forms can be administered to patients by variousroutes including, but not limited to, subcutaneous, intravenous(including bolus injection), intramuscular, and intraarterial. Becausetheir administration typically bypasses patients' natural defensesagainst contaminants, parenteral dosage forms are preferably sterile orcapable of being sterilized prior to administration to a patient.Examples of parenteral dosage forms include, but are not limited to,solutions ready for injection, dry products ready to be dissolved orsuspended in a pharmaceutically acceptable vehicle for injection,suspensions ready for injection, and emulsions.

Suitable vehicles that can be used to provide parenteral dosage formsprovided herein are well known to those skilled in the art. Examplesinclude, but are not limited to: Water for Injection USP; aqueousvehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

Compounds that increase the solubility of one or more of the activeingredients disclosed herein can also be incorporated into theparenteral dosage forms provided herein. For example, cyclodextrin andits derivatives can be used to increase the solubility of animmunomodulatory compound provided herein and its derivatives. See,e.g., U.S. Pat. No. 5,134,127, which is incorporated herein byreference.

4.6.4 Topical and Mucosal Dosage Forms

Topical and mucosal dosage forms provided herein include, but are notlimited to, sprays, aerosols, solutions, emulsions, suspensions, eyedrops or other ophthalmic preparations, or other forms known to one ofskill in the art. See, e.g., Remington's Pharmaceutical Sciences, 16thand 18th eds., Mack Publishing, Easton Pa. (1980 & 1990); andIntroduction to Pharmaceutical Dosage Forms, 4th ed., Lea & Febiger,Philadelphia (1985). Dosage forms suitable for treating mucosal tissueswithin the oral cavity can be formulated as mouthwashes or as oral gels.

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide topical and mucosal dosage forms encompassedby this disclosure are well known to those skilled in the pharmaceuticalarts, and depend on the particular tissue to which a givenpharmaceutical composition or dosage form will be applied. With thatfact in mind, typical excipients include, but are not limited to, water,acetone, ethanol, ethylene glycol, propylene glycol, butane-1,3-diol,isopropyl myristate, isopropyl palmitate, mineral oil, and mixturesthereof to form solutions, emulsions or gels, which are non-toxic andpharmaceutically acceptable. Moisturizers or humectants can also beadded to pharmaceutical compositions and dosage forms if desired.Examples of such additional ingredients are well known in the art. See,e.g., Remington's Pharmaceutical Sciences, 16th and 18th eds., MackPublishing, Easton Pa. (1980 & 1990).

The pH of a pharmaceutical composition or dosage form may also beadjusted to improve delivery of one or more active ingredients.Similarly, the polarity of a solvent carrier, its ionic strength, ortonicity can be adjusted to improve delivery. Compounds such asstearates can also be added to pharmaceutical compositions or dosageforms to advantageously alter the hydrophilicity or lipophilicity of oneor more active ingredients so as to improve delivery. In this regard,stearates can serve as a lipid vehicle for the formulation, as anemulsifying agent or surfactant, and as a delivery-enhancing orpenetration-enhancing agent. Different salts, hydrates or solvates ofthe active ingredients can be used to further adjust the properties ofthe resulting composition.

4.7 Kits

The compounds provided herein can also be provided as an article ofmanufacture using packaging materials well known to those of skill inthe art. See, e.g., U.S. Pat. Nos. 5,323,907; 5,052,558; and 5,033,252.Examples of pharmaceutical packaging materials include, but are notlimited to, blister packs, bottles, tubes, inhalers, pumps, bags, vials,containers, syringes, and any packaging material suitable for a selectedformulation and intended mode of administration and treatment.

Provided herein also are kits which, when used by the medicalpractitioner, can simplify the administration of appropriate amounts ofactive ingredients to a subject. In certain embodiments, the kitprovided herein includes a container and a dosage form of a compoundprovided herein, including a single enantiomer, a mixture of anenantiomeric pair, an individual diastereomer, or a mixture ofdiastereomers thereof; or a pharmaceutically acceptable salt, solvate,or prodrug thereof.

In certain embodiments, the kit includes a container comprising a dosageform of the compound provided herein, including a single enantiomer, amixture of an enantiomeric pair, an individual diastereomer, or amixture of diastereomers thereof; or a pharmaceutically acceptable salt,solvate, or prodrug thereof, in a container comprising one or more othertherapeutic agent(s) described herein. Kits provided herein can furthercomprise additional active ingredients such as oblimersen (Genasense®),melphalan, G-CSF, GM-CSF, EPO, topotecan, dacarbazine, irinotecan,taxotere, IFN, COX-2 inhibitor, pentoxifylline, ciprofloxacin,dexamethasone, IL2, IL8, IL 18, Ara-C, vinorelbine, isotretinoin, 13cis-retinoic acid, or a pharmacologically active mutant or derivativethereof, or a combination thereof. Examples of the additional activeingredients include, but are not limited to, those disclosed herein.

Kits provided herein can further include devices that are used toadminister the active ingredients. Examples of such devices include, butare not limited to, syringes, needle-less injectors drip bags, patches,and inhalers. The kits provided herein can also include condoms foradministration of the active ingredients.

Kits provided herein can further include pharmaceutically acceptablevehicles that can be used to administer one or more active ingredients.For example, if an active ingredient is provided in a solid form thatmust be reconstituted for parenteral administration, the kit cancomprise a sealed container of a suitable vehicle in which the activeingredient can be dissolved to form a particulate-free sterile solutionthat is suitable for parenteral administration. Examples ofpharmaceutically acceptable vehicles include, but are not limited to:aqueous vehicles, including, but not limited to, Water for InjectionUSP, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection,Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection;water-miscible vehicles, including, but not limited to, ethyl alcohol,polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles,including, but not limited to, corn oil, cottonseed oil, peanut oil,sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

The disclosure will be further understood by the following non-limitingexamples.

5. EXAMPLES 5.11-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea

To a suspension of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.18 g, 0.5 mmol) in DMF (2 mL) was added CDI(81 mg, 0.5 mmol). The mixture was stirred at RT overnight.4-Pyridin-4-ylmethyl-phenylamine (92 mg, 0.5 mmol) was added to themixture and the mixture was stirred at RT for 4 hrs. Then thetemperature was elevated to 40° C. and the mixture was stirred at thistemperature for 8 hrs. The mixture was cooled to RT, added water (5 mL),and stirred for 10 min. The suspension was filtered and the solid waswashed with water (20 mL), EtOAc (20 mL), and CH₃CN (20 mL) to give theproduct as an off-white solid (100 mg, 41% yield): HPLC: Waters SymmetryC₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm, 5% gradient 100% in 5 min,CH₃CN/0.1% H₃PO₄, 4.69 min (94%); mp: 290-292° C.; ¹H NMR (DMSO-d₆) δ1.93-2.06 (m, 1H, CHH), 2.29-2.44 (m, 1H, CHH), 2.54-2.68 (m, 1H, CHH),2.82-3.02 (m, 1H, CHH), 3.88 (s, 2H, CH₂), 4.24-4.54 (m, 4H, CH₂, CH₂),5.10 (dd, J=5.0, 13.7 Hz, 1H, NCH), 6.70 (s, 1H, NH), 7.10 (d, J=8.3 Hz,2H, Ar), 7.22 (d, J=5.3 Hz, 2H, Ar), 7.34 (d, J=8.3 Hz, 2H, Ar),7.39-7.48 (m, 1H, Ar), 7.51 (s, 1H, Ar), 7.69 (d, J=7.7 Hz, 1H, Ar),8.40-8.52 (m, 2H, Ar), 8.59 (s, 1H, NH), 10.98 (s, 1H, NH); ¹³C NMR(DMSO-d₆) δ 22.49, 31.20, 40.63, 42.76, 47.12, 51.56, 118.03, 121.83,122.91, 123.99, 126.86, 129.06, 130.27, 132.08, 138.75, 142.36, 144.87,149.42, 155.22, 163.87, 167.93, 170.98, 172.85; LCMS MH⁺=484; Anal.Calcd. for C₂₇H₂₅N₅O₄+1.5 H₂O: C, 63.52; H, 5.53; N, 13.72; S, 6.55.Found: C, 63.68; H, 5.24; N, 13.79.

5.21-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-hydroxymethyl-phenyl)-urea

To a suspension of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.18 g, 0.5 mmol) in DMF (2 mL) was added CDI(81 mg, 0.5 mmol). The mixture was stirred at RT overnight.4-Amino-benzyl alcohol (62 mg, 0.5 mmol) was added to the mixture andthe mixture was stirred at RT for 4 hrs. Then the temperature waselevated to 40° C. and the mixture was stirred at this temperature for 8hrs. The mixture was cooled to RT, added water (5 mL) and stirred for 10min. The suspension was filtered and the solid was washed with water (20mL), EtOAc (20 mL), and CH₃CN (20 mL) to give a reddish solid (100 mg,41% yield): The solid was purified on ISCO silica gel column usingmethanol and DCM as eluent to give the product as a white solid (30 mg,15% yield): HPLC: Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240nm, 5% gradient 100% in 5 min, CH₃CN/0.1% H₃PO₄, 4.99 min (97%); mp:309-311° C.; ¹H NMR (DMSO-d₆) δ 1.92-2.10 (m, 1H, CHH), 2.20-2.47 (m,1H, CHH), 2.54-2.69 (m, 1H, CHH), 2.80-3.07 (m, 1H, CHH), 4.19-4.54 (m,6H, CH₂, CH₂, CH₂), 4.94-5.05 (m, 1H, OH), 5.11 (dd, J=5.1, 13.2 Hz, 1H,CHN), 6.70 (t, J=6.0 Hz, 1H, NH), 7.16 (d, J=8.7 Hz, 2H, Ar), 7.28-7.40(m, 2H, Ar), 7.39-7.48 (m, 1H, Ar), 7.52 (s, 1H, Ar), 7.69 (d, J=7.7 Hz,1H, Ar), 8.57 (s, 1H, NH), 10.98 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 22.49,31.18, 42.77, 47.12, 51.56, 62.67, 117.49, 121.89, 122.93, 126.91,127.05, 130.29, 135.22, 138.99, 142.38, 144.88, 155.25, 167.95, 170.99,172.85; LCMS MH⁺=423; Anal. Calcd. for C₂₂H₂₂N₄O₅+0.4 H₂O: C, 61.50; H,5.35; N, 13.04. Found: C, 61.21; H, 5.05; N, 12.80.

5.31-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-4H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea

To a mixture of 5-(3-isocyanato-phenyl)-1-methyl-1H-pyrazole (0.22 g,1.1 mmol) and3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.40 g, 1.1 mmol) in acetonitrile (5 mL) wasadded TEA (0.31 mL, 2.2 mmol) at RT, and the mixture was kept for 22hrs. Water (25 mL) was added to the mixture and the mixture was stirredat RT for 3 hrs. The suspension was filtered and the solid was washedwith water (20 mL), EtOAc (20 mL), and water (20 mL) to give a solid.The solid was purified with preparative HPLC to give the product as awhite solid (122 mg, 24% yield): HPLC: Waters Symmetry C₁₈, 5 3.9×150mm, 1 mL/min, 240 nm, 30/70 CH₃CN/0.1% H₃PO₄, 3.34 min (99.8%); mp:260-262° C.; ¹H NMR (DMSO-d₆) δ 1.88-2.10 (m, 1H, CHH), 2.27-2.47 (m,1H, CHH), 2.55-2.66 (m, 1H, CHH), 2.80-3.02 (m, 1H, CHH), 3.84 (s, 3H,CH₃), 4.30 (d, J=17.6 Hz, 1H, CHH), 4.40-4.50 (m, 3H, CHH, CH₂), 5.11(dd, J=5.2, 13.3 Hz, 1H, NCH), 6.35 (d, J=1.9 Hz, 1H, Ar), 6.83 (t,J=6.1 Hz, 1H, NH), 7.06 (dt, J=1.4, 7.6 Hz, 1H, Ar), 7.31-7.38 (m, 1H,Ar), 7.40-7.48 (m, 3H, Ar), 7.53 (d, J=0.4 Hz, 1H, Ar), 7.63 (t, J=1.9Hz, 1H, Ar), 7.70 (d, J=7.9 Hz, 1H, Ar), 8.82 (s, 1H, NH), 10.98 (br.s., 1H, NH); NMR (DMSO-d₆) δ 22.48, 31.19, 37.45, 42.78, 47.11, 51.55,105.55, 117.62 (2 carbons by HMQC), 121.18, 121.87, 122.93, 126.87,129.08, 130.29, 130.51, 137.86, 140.73, 142.38, 142.82, 144.76, 155.21,167.94, 170.98, 172.84; LCMS MH⁺=473; Anal. Calcd. for C₂₅H₂₄N₆O₄: C,63.55; H, 5.12; N, 17.79. Found: C, 63.36; H, 5.17; N, 17.72.

5.41-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-4H-isoindol-5-ylmethyl]-3-[3-(2-methyl-imidazol-1-yl)-phenyl]-urea;formic acid

A mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.40 g, 1.1 mmol) and CDI (0.19 g, 1.2 mmol)in DMF (5 mL) was stirred at RT for 18 hrs. To the mixture was added3-(2-methyl-imidazol-1-yl)-phenylamine (0.19 g, 1.1 mmol) at RT, and themixture was stirred at 60° C. for 24 hrs. To the mixture was added water(25 mL) and ether (20 mL). The mixture was stirred at RT for 2 hrs. Thesuspension was filtered and the solid was washed with water (20 mL),ethyl acetate (20 mL), and water (20 mL) to give a solid. The solid waspurified with preparative HPLC to give the product as a white solid (100mg, 20% yield): HPLC: Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 mL/min,240 nm, 5% gradient 95% in 5 min, CH₃CN/0.1% H₁PO₄, 4.48 min (96.8%);mp: 218-220° C.; ¹H NMR (DMSO-d₆) δ 1.89-2.12 (m, 1H, CHH), 2.25-2.29(m, 3H, CH₃), 2.30-2.46 (m, 1H, CHH), 2.54-2.68 (m, 1H, CHH), 2.78-3.04(m, 1H, CHH), 4.31 (d, J=17.4 Hz, 1H, CHH), 4.38-4.53 (m, 3H, CHH, CH₂),5.11 (dd, J=5.1, 13.2 Hz, 1H, NCH), 6.89 (d, J=1.3 Hz, 1H. Ar),6.92-7.05 (m, 2H, Ar, NH), 7.23 (d, J=1.3 Hz, 1H, Ar), 7.32-7.40 (m, 2H,Ar), 7.41-7.49 (m, 1H, Ar), 7.52 (s, 1H, Ar), 7.57-7.65 (m, 1H, Ar),7.69 (d, J=7.9 Hz, 1H, Ar), 8.18 (s, 1H, HCOOH), 9.03 (s, 1H, NH), 10.98(s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 13.60, 22.49, 31.20, 42.77, 47.12,51.58, 114.22, 116.89, 117.61, 120.68, 121.88, 122.93, 126.89, 127.13,129.63, 130.30, 137.90, 141.50, 142.38, 143.43, 144.69, 155.16, 163.44,167.93, 170.98, 172.85; LCMS MH⁺=473; Anal. Calcd. forC₂₅H₂₄N₆O₄+HCOOH+1.5 H₂O: C, 57.24; H, 5.36; N, 15.40. Found: C, 57.43;H, 5.11; N, 15.57.

5.51-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[4-(4-methyl-4H-[1,2,4]-triazol-3-yl)-phenyl]-urea

To a stirred suspension of4-(4-methyl-4H-[1,2,4]triazol-3-yl)-phenylamine (0.31 g, 1.77 mmol) inDMF (10 mL) at 40° C. was added CDI (0.32 g, 1.94 mmol). The mixture wasstirred for 15 min, followed by addition of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.65 g, 1.77 mmol). Heating was stopped after1.5 hrs and the mixture was stirred at RT overnight. Solvent wasevaporated and the residue was purified by preparative HPLC to give theproduct as a white solid (0.13 g, 15% yield): HPLC, Waters Symmetry C₁₈,5 μm, 3.9×150 mm, 1 ml/min, 240 nm, 14/86 CH₃CN/0.1% H₁PO₄, 4.70 min(93.9%); mp, 248-250° C.; ¹H NMR (DMSO-d₆) δ 1.93-2.06 (m, 1H, CHH),2.29-2.46 (m, 1H, CHH), 2.55-2.70 (m, 1H, CHH), 2.82-3.02 (m, 1H, CHH),3.72 (s, 3H, CH₃), 4.24-4.55 (m, 4H, CH₂, CH₂), 5.11 (dd, 4.9, 13.2 Hz,1H, NCH), 6.95 (t, J=5.7 Hz, 1H, NH), 7.41-7.81 (m, 7H, ArH), 8.51 (s,1H, ArH), 9.02 (s, 1H, NH), 10.99 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ22.49, 31.20, 31.94, 42.79, 47.13, 51.58, 117.50, 119.49, 121.88,122.94, 126.91, 128.80, 130.32, 141.84, 142.39, 144.72, 145.76, 153.08,155.12, 167.95, 170.99, 172.85; LC/MS MH⁺=474; Anal. Calcd. ForC₂₄H₂₃N₇O₄: C, 60.88; H, 4.90; N, 20.71. Found: C, 58.28; H, 4.67; N,19.49 (Note: This analysis was off, and both HPLC and ¹H NMR showed 6%impurity).

5.61-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea

To a mixture of 4-(3-isocyanato-phenyl)-2-methyl-thiazole (0.25 g, 1.2mmol) and3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.43 g, 1.2 mmol) in acetonitrile (5 mL) wasadded TEA (0.33 mL, 2.3 mmol) at RT, and the mixture was kept for 3 hrs.Water (25 mL) was added to the mixture and the mixture was stirred at RTfor 3 hrs. The suspension was filtered and the solid was washed withwater (20 mL), ethyl acetate (20 mL), and water (20 mL) to give a solid.The solid was purified with preparative HPLC to give the product as awhite solid (160 mg, 28% yield): HPLC: Waters Symmetry C₁₈, 5 μm,3.9×150 mm, 1 mL/min, 240 nm, 30/70 CH₃CN/0.1% H₁PO₄, 5.93 min (99.4%);mp: 252-254° C.; ¹H NMR (DMSO-d₆) δ 1.92-2.06 (m, 1H, CHH), 2.27-2.47(m, 1H, CHH), 2.54-2.66 (m, 1H, CHH), 2.71 (s, 3H, CH₃), 2.81-3.01 (m,1H, CHH), 4.31 (d, J=17.6 Hz, 1H, CHH), 4.38-4.53 (m, 3H, CHH, CH₂),5.11 (dd, J=5.1, 13.2 Hz, 1H, NCH), 6.74 (t, J=6.0 Hz, 1H, NH),7.20-7.32 (m, 1H, Ar), 7.40 (ddd, J=1.1, 2.3, 8.1 Hz, 1H, Ar), 7.46 (dq,J=1.4, 7.6 Hz, 2H, Ar), 7.53 (s, 1H, Ar), 7.70 (d, J=7.9 Hz, 1H, Ar),7.81 (s, 1H, Ar), 8.02 (t, J=1.9 Hz, 1H, Ar), 8.79 (s, 1H, NH), 10.98(s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 18.91, 22.51, 31.18, 42.79, 47.12,51.56, 113.55, 115.55, 117.28, 118.86, 121.86, 122.94, 126.89, 129.01,130.29, 134.58, 140.82, 142.39, 144.90, 153.89, 155.21, 165.29, 167.95,170.99, 172.85; LCMS MH⁺=490; Anal. Calcd. for C₂₅H₂₃N₅O₄S: C, 61.34; H,4.74; N, 14.31; S, 6.55. Found: C, 61.09; H, 4.60; N, 14.19; S, 6.49.

5.71-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(1-methyl-M-pyrazol-3-yl)-phenyl]-urea

To a stirred suspension of 3-(3-isocyanato-phenyl)-1-methyl-1H-pyrazole(0.25 g, 1.25 mmol) and3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.46 g, 1.25 mmol) in acetonitrile (5 mL) atRT was added TEA (0.35 mL, 2.51 mmol). The mixture was stirred for 4hrs, followed by addition of 1N HCl (10 mL), which was stirred for 10min. The mixture was purified by preparative HPLC to give the product asan off-white solid (0.22 g, 38% yield): HPLC, Waters Symmetry C₁₈, 5 μm,3.9×150 mm, 1 ml/min, 240 nm, 25/75 CH₃CN/0.1% H₃PO₄, 5.99 min (99.9%);mp, 232-234° C.; ¹H NMR (DMSO-d₆) δ 1.93-2.07 (m, 1H, CHH), 2.29-2.46(m, 1H, CHH), 2.55-2.68 (m, 1H, CHH), 2.82-3.01 (m, 1H, CHH), 3.87 (s,3H, CHH), 4.24-4.55 (m, 4H, CH₂, CH₂), 5.11 (dd. J=5.0, 13.3 Hz, 1H,NCH), 6.57 (d, J=2.3 Hz, 1H, ArH), 6.73 (t, J=5.9 Hz, 1H, NH), 7.18-7.37(m, 3H, ArH), 7.46 (d, J=7.9 Hz, 1H, ArH), 7.53 (s, 1H, ArH), 7.65-7.77(m, 2H, ArH), 7.89 (s, 1H, ArH), 8.71 (s, 1H, NH), 10.98 (s, 1H, NH);¹³C NMR (DMSO-d₆) δ 22.51, 31.18, 38.60, 42.79, 47.13, 51.58, 102.33,114.31, 116.77, 118.16, 121.86, 122.93, 126.89, 128.85, 130.29, 132.17,133.85, 140.66, 142.39, 144.91, 150.03, 155.22, 167.95, 170.99, 172.85;LC/MS MH⁺=473; Anal. Calcd. For C₂₅H₂₄N₆O₄+0.5 H₂O: C, 62.36; H, 5.23;N, 17.45. Found: C, 62.06; H, 5.19; N, 17.28.

5.81-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)ureaformate

To a stirred mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionehydrochloride (0.37 g, 1.00 mmol) and 4-(3-isocyanatobenzyl)-morpholine(0.22 g, 1.00 mmol) in acetonitrile (10 mL) was added TEA (0.28 mL, 2.00mmol) at RT under nitrogen. After 2 hrs, additional4-(3-isocyanatobenzyl)morpholine (0.22 g, 1.00 mmol) and TEA (0.28 mL,2.00 mmol) were added. After 12 hrs, undesired solid was filtered andfiltrate was concentrated. The residue was dissolved in acetonitrile andwas purified by preparative HPLC (gradient: CH₃CN+0.1% formicacid/H₂O+0.1% formic acid: 10/90 for 5 min, to 100/0 in 10 min, 100/0for 5 min). After evaporation of the solvent, the residue was trituratedin ether (20 mL) for 1 hr. The product was then isolated by filtrationand dried in vacuo to give the product as a beige solid (0.16 g, 30%yield): HPLC: Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm,gradient: CH₃CN/0.1% H₃PO₄: 10/90 to 90/10 in 10 min, 90/10 (5 min):4.64 min (95.84%); mp: 198-200° C.; ¹H NMR (DMSO-d₆) δ 1.83-2.12 (m, 1H,CHH), 2.21-2.47 (m, 5H, CHH, CH₂, CH₂), 2.54-2.69 (m, J=11.0 Hz, 1H,CHH), 2.79-3.03 (m, 1H, CHH), 3.38 (s, 2H, CH₂), 3.52-3.72 (m, 4H, CH₂,CH₂), 4.31 (d, J=17.2 Hz, 1H, CHH), 4.37-4.55 (m, 3H, CHH, CH₂), 5.11(dd, J=5.1, 13.2 Hz, 1H, CH), 6.83 (d, J=7.6 Hz, 1H, Ar), 6.99 (t, J=5.6Hz, 1H, NH), 7.15 (t, J=7.7 Hz, 1H, Ar), 7.32 (d, J=8.3 Hz, 1H, Ar),7.39 (s, 1H, Ar), 7.44 (d, J=7.7 Hz, 1H, Ar), 7.52 (s, 1H. Ar), 7.69 (d,J=7.7 Hz, 1H, Ar), 8.29 (br. s., 1H, HCOO), 8.90 (s, 1H, NH), 10.98 (br.s., 1H, NH); ¹³C NMR (DMSO-d₆) δ 22.51, 31.20, 42.73, 47.12, 51.56,53.20, 62.69, 66.17, 116.42, 118.13, 121.72, 121.83, 122.90, 126.86,128.34, 130.25, 138.24, 140.48, 142.36, 144.99, 155.31, 164.30, 167.95,170.98, 172.85; LCMS: MH⁺=492; Anal. Calcd. for C₂₇H₃₁N₅O₇+3 H₂O: C,54.82; H, 6.30; N, 11.84. Found: C, 55.12; H, 6.12; N, 11.72.

5.91-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea

To a stirred mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionehydrochloride (1.00 g, 2.70 mmol) and 4-methyl-3-nitrophenylisocyanate(0.48 mL, 2.70 mmol) in acetonitrile (20 mL) was added TEA (0.75 mL,5.40 mmol) at RT under nitrogen. After 2 hrs, 1N HCl (20 mL) was addedand the solids were isolated by filtration and washed with water (3×20mL). The crude product was triturated in EtOAc (50 mL) for 12 hrs. Theproduct was isolated by filtration and dried in vacuo to give theproduct as a yellow solid (0.74 g, 61% yield): HPLC: Waters SymmetryC₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm, gradient: CH₃CN/0.1% H₃PO₄:10/90 to 90/10 in 10 min, 90/10 (5 min): 7.94 min (96.79%); mp: 230-232°C.; ¹H NMR (DMSO-d₆) δ 1.87-2.08 (m, 1H, CHH), 2.26-2.48 (m, 3H, CHH,CH₃), 2.60 (d, J=17.6 Hz, 1H, CHH), 2.79-3.07 (m, 1H, CHH), 4.31 (d,J=17.4 Hz, 1H, CHH), 4.38-4.55 (m, 3H, CHH, CH₂), 5.11 (dd, J=5.0, 13.3Hz, 1H, CH), 6.92 (t, J=5.9 Hz, 1H, NH), 7.34 (d, J=8.3 Hz, 1H, Ar),7.45 (d, J=7.9 Hz, 1H, Ar), 7.49-7.60 (m, 2H, Ar), 7.70 (d, J=7.7 Hz,1H, Ar), 8.26 (d, J=2.3 Hz, 1H, Ar), 9.07 (s, 1H, NH), 10.99 (s, 1H,NH); ¹³C NMR (DMSO-d₆) 18.95, 22.49, 31.20, 42.82, 47.12, 51.56, 112.74,121.92, 122.45, 122.94, 124.71, 126.92, 130.33, 132.84, 139.46, 142.38,144.59, 148.72, 155.02, 167.93, 170.98, 172.85; LCMS: MH⁺=452; Anal.Calcd. for C₂₂H₂₁N₄O₆: C, 58.53; H, 4.69; N, 15.51. Found: C, 58.23; H,4.58; N, 15.34.

5.101-(3-amino-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea

To a solution of1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea(0.30 g, 0.66 mmol) in DMF (50 mL) was added Pd—C (0.10 g, 10% weight).The reaction mixture was hydrogenated with a Parr-shaker at 55 psi.After 16 hrs. the mixture was filtered through a celite pad, which waswashed with additional DMF (20 mL). The filtrate was then evaporated andthe residue was stirred in water (100 mL) for 3 hrs. The solid wasfiltered, washed with additional water (50 mL), and dried. The crudegreen product was dissolved in DMF (100 mL), decolorizing carbon wasadded, and the reaction mixture was stirred for 3 hrs. The mixture wasthen filtered through a celite pad, which was washed with additional DMF(50 mL). The filtrate was then evaporated and the residue was stirred inwater (100 mL) for 4 hrs. The solid was filtered, washed with additionalwater (50 mL), and dried in vacuo. The solid was triturated with etherfor 1 hr and the product was isolated by filtration to give the productas a pale green solid (0.22 g, 79% yield): HPLC: Waters Symmetry C₁₈, 5μm, 3.9×150 mm, 1 mL/min, 240 nm, gradient: CH₃CN/0.1% H₁PO₄: 10/90 to90/10 in 10 min, 90/10 (5 min): 4.50 min (96.74%); mp: 228-230° C.; ¹HNMR (DMSO-d₆) δ 1.82-2.10 (m, 4H, CHH, CH₃), 2.25-2.47 (m, 1H, CHH),2.59 (d. J=18.3 Hz, 1H, CHH), 2.77-3.02 (m, 1H, CHH), 4.30 (d, J=17.6Hz, 1H, CHH), 4.35-4.53 (m, 3H, CH₂, CHH), 4.93 (br. s., 2H, NH₂), 5.11(dd, J=5.0, 12.9 Hz, 1H, CH), 6.51 (dd, J=1.9, 7.9 Hz, 1H, Ar), 6.58 (t,J=5.7 Hz, 1H, NH), 6.75 (d, J=8.5 Hz, 2H, Ar), 7.43 (d, J=7.9 Hz, 1H,Ar), 7.50 (s, 1H, Ar), 7.69 (d, J=7.7 Hz, 1H. Ar), 8.23 (s, 1H, NH),10.98 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 16.75, 22.49, 31.20, 42.74,47.10, 51.56, 104.16, 106.57, 114.59, 121.86, 122.90, 126.89, 129.79,130.25, 138.75, 142.36, 145.06, 146.12, 155.22, 167.95, 170.98, 172.85;LCMS: MH⁺=422; Anal. Calcd. for C₂₂H₂₃N₅O₄+0.1 H₂O+0.5 Et₂O: C, 62.62;H, 6.17; N, 15.21. Found: C, 62.30; H, 5.89; N, 14.89.

5.111-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea

TEA (0.20 g, 2.00 mmol) was added to a stirred mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonate (0.37 g, 1.00 mmol) and 1-isocyanato-3-phenoxy-benzene(0.212 g, 1.00 mmol) in acetonitrile (10 mL) under nitrogen at RT. After3 hrs, 1N HCl (10 mL) was added, and the mixture was stirred for 10 min.The solids were isolated by filtration, and washed with water (20 mL)and acetonitrile (10 mL). The crude product was dissolved in a minimalamount of DMF and the product was precipitated by slow addition of water(˜30 mL). The solids were collected by filtration, washed with Et₂O, anddried in vacuo for 18 hrs to give the product as a white solid (422 mg,87%): HPLC: Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 ml/min, 240 nm,35/65, CH₃CN/0.1% H₁₁PO₄, 3.76 min (99.3%); mp: 224-226° C.; NMR(DMSO-d₆) δ 1.99 (s, 4H, CH, CH₃), 2.29-2.46 (m, 1H, CH), 2.55-2.66 (m,1H, CH), 2.80-3.01 (m, 1H, CH), 4.18-4.56 (m, 4H, CH₂, CH₂), 5.10 (dd.J=4.9, 13.2 Hz, 1H, CH), 6.71 (t, J=5.9 Hz, 1H, NH), 7.23-7.36 (m, 1H,Ar), 7.36-7.48 (m, 2H, Ar), 7.51 (s, 1H, Ar), 7.69 (d, J=7.7 Hz, 1H,Ar), 8.55 (s, 1H, NH), 9.75 (s, 1H, NH), 10.67-11.27 (m, 1H, NH); ¹¹CNMR (DMSO-d₆) δ 22.49, 23.82, 31.18, 42.77, 47.12, 51.56, 118.12,119.57, 121.85, 122.91, 126.88, 130.26, 133.18, 135.70, 142.36, 144.94,155.29, 167.67, 167.95, 170.98, 172.85; LCMS: MH⁺=485; Anal. Calcd. forC₂₇H₂₄N₄O₅: C, 66.93; H, 4.99; N, 11.56. Found: C, 67.03; H, 4.72; N,11.41.

5.121-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea

TEA (0.20 g, 2.0 mmol) was added to a stirred mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonate (0.37 g, 1.0 mmol) and 1-isocyanato-4-nitrobenzene (164mg, 1.0 mmol) in acetonitrile (10 mL) under nitrogen. The mixturestirred at RT for 3 hrs, during which time it remained as a suspension.A 1N HCl solution (10 mL) was added, and the mixture was stirred for 10min. The solid was isolated by filtration and washed with additionalwater (20 mL) and acetonitrile (10 mL). The solid was dissolved inminimal amount of DMF and the product was precipitated by slow additionof water (˜30 mL). The solid was collected by filtration and washed withEt₂O to remove most of the residual yellow color. The remaining solidwas dried in a vacuum oven overnight to provide the product as anoff-white solid (330 mg, 75%): HPLC: Waters Symmetry C₁₈, 5 μm, 3.9×150mm, 1 ml/min, 240 nm, 30/70, CH₁CN/0.1% H₃PO₄, 6.15 min (97.1%); mp:272-274° C.; ¹H NMR (DMSO-d₆) δ 1.84-2.13 (m, 1H, CHH), 2.24-2.44 (m,1H, CHH), 2.55-2.66 (m, 1H, CHH), 2.77-3.02 (m, 1H, CHH), 4.16-4.55 (m,4H, CH₂, CH₂), 5.11 (dd, J=4.7, 13.0 Hz, 1H, CH), 7.07 (t, J=5.6 Hz, 1H,NH), 7.37-7.83 (m, 5H, Ar), 8.15 (d, J=8.9 Hz, 2H, Ar), 9.48 (s, 1H,NH), 10.98 (s, 1H, NH); ¹³C NMR (DMSO-d₆) d 22.48, 31.17, 42.83, 47.11,51.57, 116.97, 121.94, 122.96, 125.09, 126.93, 130.38, 140.48, 142.41,144.26, 147.04, 154.51, 167.91, 170.98, 172.85; LCMS: MH⁺=438; Anal.Calcd. for C₂₁H₁₉N₅O₆+0.5 H₂O: C, 56.50; H, 4.52; N, 15.69. Found: C,56.45; H, 4.31; N, 15.71.

5.13N-(4-{3-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-phenyl)-acetamide

Step 1: Preparation of1-(4-amino-phenyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea

To a stirred mixture of14(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea(150 mg, 0.343 mmol) in EtOH (2 mL) was added sodium dithionite (597 mg,3.43 mmol) in water (2 mL). The resulting mixture was heated to 60° C.for 20 min at which time LC-MS indicated complete disappearance of nitrostarting material. The reaction mixture was combined with the crudeproduct from a separate run and concentrated in vacuo. The residue wasdissolved in minimal DMF and chromatographed on a C-18 preparative HPLCcolumn equipped with mass triggered collection. The desired fractionswere combined and concentrated in vacuo to provide1-(4-amino-phenyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureaas a pale yellow solid (90 mg, 40% combined average yield from twoseparate runs): mp: >400° C.; LCMS: MH⁺=408.

Step 2: Preparation ofN-(4-{3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-phenyl)-acetamide

1-(4-Amino-phenyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea(64 mg, 0.157 mmol) was stirred at room temperature in acetic anhydride(5 mL) for 2 hrs. The volatiles were removed in vacuo and the residuewas dissolved in minimal DMF and purified on a C-18 preparative HPLCcolumn. The desired fractions were combined and concentrated in vacuo toprovide the product as a pale yellow solid (37 mg, 52% yield): HPLC:Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 ml/min, 240 nm, 17/83,CH₃CN/0.1% H₃PO₄, 5.19 min (97.4%); mp: 265-267° C.; ¹H NMR (DMSO-d₆) δ1.99 (s, 4H, CH, CH₃), 2.29-2.46 (m, 1H, CH), 2.55-2.66 (m, 1H, CH),2.80-3.01 (m, 1H, CH), 4.18-4.56 (m, 4H, CH₂, CH₂), 5.10 (dd, J=4.9,13.2 Hz, 1H, CH), 6.71 (t, J=5.9 Hz, 1H, NH), 7.23-7.36 (m, 1H, Ar),7.36-7.48 (m, 2H, Ar), 7.51 (s, 1H, Ar), 7.69 (d, J=7.7 Hz, 1H, Ar),8.55 (s, 1H, NH), 9.75 (s, 1H, NH), 10.67-11.27 (m, 1H, NH); ¹³C NMR(DMSO-d₆) δ 22.49, 23.82, 31.18, 42.77, 47.12, 51.56, 118.12, 119.57,121.85, 122.91, 126.88, 130.26, 133.18, 135.70, 142.36, 144.94, 155.29,167.67, 167.95, 170.98, 172.85; LCMS: MH⁺=450; Anal. Calcd. forC₂₃H₂₃N₅O₅+1.0 H₂O: C, 59.09; H, 5.39; N, 14.98. Found: C, 58.75; H,4.99; N, 14.59.

5.14 3-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dhydro-1H-isoindol-5-ylmethyl]-1-methyl-1-phenyl-urea

To a suspension of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.18 g, 0.5 mmol) in CH₃CN (10 mL) was addedDIPEA (0.4 mL, 2.5 mmol) and N-methylphenyl carbamic chloride (178 mg,1.05 mmol). The mixture was stirred at RT overnight. The suspension wasfiltered and the solid was washed with water (20 mL), ethyl acetate (20mL), and CH₃CN (20 mL) to give the product as a white solid (200 mg, 47%yield): HPLC: Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm,30/70 CH₃CN/0.1% H₃PO₄, 3.56 min (96%); mp: 168-170° C.; ¹H NMR(DMSO-d₆) δ 1.89-2.07 (m, 1H, CHH), 2.30-2.47 (m, 1H, CHH), 2.55-2.69(m, 1H, CHH), 2.79-3.01 (m, 1H, CHH), 3.18 (s, 3H, CH₃), 4.22-4.58 (m,4H, CH₂, CH₂), 5.11 (dd, J=5.0, 13.3 Hz, 1H, NCH), 6.68 (t, J=5.9 Hz,1H, NH), 7.16-7.35 (m, 3H, Ar), 7.36-7.44 (m, 3H, Ar), 7.46 (s, 1H, Ar),7.66 (d, J=7.7 Hz, 1H, Ar), 10.98 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ22.51, 31.21, 37.14, 43.76, 47.10, 51.55, 121.85, 122.71, 125.78,126.54, 126.86, 129.23, 130.04, 142.19, 144.02, 145.41, 156.74, 168.02,171.02, 172.86; LCMS MH⁺=407; Anal. Calcd. for C₂₁H₁₂N₄O₄+0.5 H₂O: C,63.60; H, 5.58; N, 13.49; S, 6.55. Found: C, 63.61; H, 5.51; N, 13.48.

5.151-Biphenyl-4-yl-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea

To a suspension of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.37 g, 1 mmol) in DMF (2 mL) was added CDI(162 mg, 1 mmol). The mixture was stirred at RT overnight.4-Phenylaniline (169 mg, 1 mmol) was added to the mixture and themixture was stirred at RT for 4 hrs. Then the temperature was elevatedto 40° C. and the mixture was stirred at this temperature for 8 hrs. Themixture was cooled to RT. The suspension was filtered and the filtratewas added CH₃CN (5 mL) and the resulted suspension was filtered. Thecollected solid was recrystallized from DMF to give the product as awhite solid (30 mg, 15% yield): HPLC: Waters Symmetry C₁₈, 5 μm, 3.9×150mm, 1 mL/min, 240 nm, 50/50 CH₃CN/0.1% H₃PO₄, 2.78 min (95%); ¹H NMR(DMSO-d₆) δ 2.02 (br. s., 1H, CHH), 2.28-2.47 (m, 1H, CHH), 2.60 (d,J=18.5 Hz, 1H, CHH), 2.82-3.04 (m, 1H, CHH), 4.23-4.59 (m, 4H, CH₂,CH₂), 5.11 (dd, J=4.9, 13.2 Hz, 1H, NCH), 6.73-6.83 (m, 1H, NH),7.22-7.35 (m, 1H, Ar), 7.37-7.66 (m, 10H, Ar), 7.70 (d, J=7.9 Hz, 1H,Ar), 8.75 (s, 1H, NH), 10.98 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 22.51,31.20, 42.80, 47.13, 51.58, 118.09, 121.89, 122.94, 125.99, 126.64,126.85, 128.82, 130.30, 132.86, 139.93, 142.40, 144.81, 155.18, 167.95,170.99, 172.85; LCMS MH⁺=423.

5.161-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(3-methyl-[1,2,4]oxadiazol-5-yl)-phenyl]-urea

To a stirred suspension of3-(3-methyl-[1,2,4]oxadiazol-5-yl)-phenylamine (0.22 g, 1.09 mmol) and3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.40 g, 1.09 mmol) in acetonitrile (5 mL) atRT was added TEA (0.31 mL, 2.19 mmol). The mixture was stirred for 3hrs, followed by addition of 1N HCl (10 mL), which was stirred for 10min. The mixture was purified by preparative HPLC to give the product asa white solid (0.10 g, 19% yield): HPLC, Waters Symmetry C₁₈, 5 μm,3.9×150 mm, 1 ml/min, 240 nm, 30/70 CH₃CN/0.1% H₃PO₄, 4.37 min (99.3%);mp, 242-244° C.; ¹H NMR (DMSO-d₆) δ 1.92-2.07 (m, 1H, CHH), 2.30-2.46(m, 4H, CH₃, CHH), 2.55-2.67 (m, 1H, CHH), 2.82-3.02 (m, 1H, CHH),4.24-4.56 (m, 4H, CH₁, CH₂), 5.11 (dd, J=5.1, 13.2 Hz, 1H, NCH), 6.91(t, J=5.8 Hz, 1H, NH), 7.39-7.80 (m, 6H, ArH), 8.37 (s, 1H, ArH), 9.05(s, 1H, NH), 10.98 (s, 1H, NH); ¹³C NMR (DMSO-d₆) 11.25, 22.49, 31.20,42.83, 47.12, 51.58, 116.37, 120.24, 121.93, 121.96, 122.94, 123.77,126.94, 129.90, 130.33, 141.41, 142.39, 144.68, 155.10, 167.61, 167.93,170.99, 172.85, 174.86. LC/MS MH=475; Anal. Calcd. For C₂₄H₂₂N₆O₅+0.4H₂O: C, 59.85; H, 4.77; N, 17.45. Found: C, 59.53; H, 4.68; N, 17.30.

5.171-(3-Aminophenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea

Step 1

To a stirred mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionehydrochloride (0.74 g, 2.00 mmol) and 3-nitropheny isocyanate (0.33 g,2.00 mmol) in acetonitrile (20 mL) was added TEA (0.56 mL, 4.00 mmol) atRT under nitrogen. After 12 hrs, the solid was filtered and purified bypreparative HPLC (gradient: CH₃CN/H₂O: 15/85 for 5 min, to 100/0 in 10min, 100/0 for 5 min). After evaporation of the solvent, the residue wastriturated in ether (20 mL) for 1 hr. The product was then isolated byfiltration and dried in vacuo to give1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-nitrophenyl)ureaas a yellow solid (0.34 g, 39% yield).

Step 2

To a solution of1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-nitrophenyl)urea(0.33 g, 0.80 mmol) in DMF (80 mL) was added Pd—C (0.10 g, <10% weight).The reaction mixture was hydrogenated with a Parr-shaker at 55 psi.After 12 hrs, the mixture was filtered through a celite pad, which waswashed with additional DMF (50 mL). The filtrate was then evaporated andthe residue was stirred in water (150 mL) for 3 hrs. The solid wasfiltered, washed with additional water (50 mL), and dried. The crudeproduct was dissolved in DMF (50 mL), decolorizing carbon was added, andthe reaction mixture was stirred for 12 hrs. The mixture was thenfiltered through a celite pad, which was washed with additional DMF (50mL). The filtrate was then evaporated and the residue was stirred inwater (100 mL) for 3 hrs. The solid was filtered, washed with additionalwater (50 mL) and dried in vacuo to give the product as a pale yellowsolid (0.24 g, 77% yield): HPLC: X-Terra RP 18, 3.9×150 mm, 5 μm, 1mL/min, 240 nm: CH₃CN/0.1% (HCO₂)NH₄: 15/85: 7.95 min (95.27%); mp:233-235° C.; NMR (DMSO-d₆) δ 1.78-2.08 (m, 1H, CHH), 2.38 (qd, J=4.5,13.2 Hz, 1H, CHH), 2.59 (d, J=17.8 Hz, 1H, CHH), 2.79-3.06 (m, 1H, CHH),4.18-4.36 (m, 1H, CHH), 4.36-4.60 (m, 3H, CHH, CH₂), 4.91-5.36 (m, 3H,NH₂, CH), 6.15 (ddd, J=0.9, 2.1, 7.9 Hz, 1H, Ar), 6.48-6.58 (m, 1H, Ar),6.63 (t, J=6.0 Hz, 1H, NH), 6.76 (t, J=2.0 Hz, 1H, Ar), 6.85 (t, J=7.9Hz, 1H, Ar), 7.44 (d, J=7.7 Hz, 1H, Ar), 7.51 (s, 1H, Ar), 7.69 (d,J=7.9 Hz, 1H, Ar), 8.32 (s, 1H, NH), 10.98 (s, 1H, NH); ¹³C NMR(DMSO-d₆) δ 22.51, 31.20, 42.73, 47.12, 51.58, 103.78, 106.20, 107.79,121.86, 122.91, 126.89, 128.89, 130.26, 140.92, 142.38, 144.99, 148.44,155.16, 167.95, 170.99, 172.85; LCMS: MH⁺=408; Anal. Calcd. forC₂₁H₂₁N₅O₄: C, 61.91, H, 5.20; N, 17.19. Found: C, 62.40, H, 5.67; N,15.59.

5.18 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydroH-isoindol-5-ylmethyl]-3-[3-(pyridin-2-yloxy)-phenyl]-urea

3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonate (1.11 g, 3.0 mmol) and 1,1′-Carbonyldiimidazole (535mg, 3.3 mmol) were suspended in dry DMF (20 mL) and the mixture wasstirred at rt for 24 h. While stirring, a portion of the reactionmixture (6.7 mL, ˜1 mmol) was transferred to a vial containing3-(Pyridin-2-yloxy)-phenylamine (205 mg, 1.1 mmol). The resultingmixture was stirred at rt overnight and the reaction progress wasmonitored by LCMS. After 48 h, additional3-(Pyridin-2-yloxy)-phenylamine (37 mg, 0.2 mmol) was transferred to thereaction mixture and stirring continued for another 24 h. The reactionmixture was acidified with acidic acid and water. The volatiles wereremoved in vacuo and the residue was dissolved in DMF and purified usingC-18 preparatory HPLC to give1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(pyridin-2-yloxy)-phenyl]-ureaas a white solid (310 mg, 64% yield): HPLC:Waters Symmetry C₁₈, 5 μm,3.9×150 mm, 1 ml/min, 240 nm, 30/70 CH₃CN/0.1% H₃PO₄, 4.81 min (98.6%);mp: 298-300° C.; ¹H NMR (DMSO-d₆) δ 1.84-2.06 (m, 1H, CHH), 2.19-2.44(m, 1H, CHH), 2.54-2.68 (m, 1H, CHH), 2.79-3.05 (m, 1H, CHH), 4.08-4.60(m, 4H, CH₂, CH₂), 5.10 (dd, J=5.1, 13.2 Hz, 1H, CH), 6.64 (dd, J=1.4,8.0 Hz, 1H, Ar), 6.81 (t, J=5.9 Hz, 1H, NH), 6.99 (d, J=8.3 Hz, 1H, Ar),7.07-7.18 (m, 2H, Ar), 7.24 (t, J=8.1 Hz, 1H, Ar), 7.33 (t, J=2.1 Hz,1H, Ar), 7.43 (d, J=7.7 Hz, 1H, Ar), 7.50 (s, 1H, Ar), 7.68 (d, J=7.7Hz, 1H, Ar), 7.78-7.97 (m, 1H, Ar), 8.16 (dd, J=1.5, 4.9 Hz, 1H, Ar),8.82 (s, 1H, NH), 10.97 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 22.49, 31.18,42.73, 47.12, 51.56, 110.23, 111.51, 113.54, 113.68, 118.99, 121.85,122.93, 126.86, 129.56, 130.29, 140.10, 141.79, 142.39, 144.77, 147.52,154.37, 155.09, 163.03, 167.93, 170.99, 172.85; LCMS: MH⁺=486; AnalCalcd for C₂₆H₂₃N₅O₅+0.3 H₂O: C, 63.61; H, 4.85; N, 14.27. Found: C,63.62; H, 4.62; N, 14.18.

5.191-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(piperidin-4-yloxy)phenyl)urea

Using the procedure as described in Section 5.15, the product isprepared from 3-(piperidin-4-yloxy)aniline and3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt.

5.201-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea

Step 1

Using the procedure as described in Section 5.15,1-(3-(tert-butyldimethylsilyloxy)-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureais prepared from 3-(tert-butyldimethylsilyloxy)-4-methylaniline and3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt.

Step 2

A mixture of1-(3-(tert-butyldimethylsilyloxy)-4-methylphenyl)-3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea(0.54 g, 1.0 mmol) and cesium fluoride (0.15 g, 1.0 mmol) in DMF (10 mL)is heated to 70° C. for 8 hrs. The mixture is cooled and diluted withwater (10 mL). The solid precipitate is filtered, rinsed with water (10mL), and dried under vacuum to provide the product.

5.215-(3-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)-2-methylphenyl2-aminoacetate hydrochloride

Step 1

Using the procedure as described in Section 5.15,5-(3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)-2-methylphenyl2-(tert-butoxycarbonylamino)acetate is prepared from5-amino-2-methylphenyl 2-(tert-butoxycarbonylamino)acetate and3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt.

Step 2

To a mixture of5-(3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)-2-methylphenyl2-(tert-butoxycarbonylamino)acetate (0.58 g, 1.0 mmol) in DCM (50 mL) isadded 2M HCl in ether (1 mL), and the mixture is stirred for 24 hrs. Thesolid precipitate is filtered, rinsed with DCM (10 mL), and dried undervacuum to provide the product.

5.225-(34(2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)-2-methylphenyl2-(piperazin-1-yl)acetate hydrochloride

Step 1

Using the procedure as described in Section 5.15, tert-butyl4-(2-(5-(3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)-2-methylphenoxy)-2-oxoethyl)piperazine-1-carboxylateis prepared from tert-butyl4-(2-(5-amino-2-methylphenoxy)-2-oxoethyl)piperazine-1-carboxylate and3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt.

Step 2

To a mixture of tert-butyl4-(2-(5-(3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)-2-methylphenoxy)-2-oxoethyl)piperazine-1-carboxylate(0.65 g, 1.0 mmol) in methylene chloride (50 mL) is added 2M HCl inether (1 mL), and the mixture will be stirred for 24 hrs. The solidprecipitate is filtered, rinsed with DCM (10 mL), and dried under vacuumto provide the product.

5.231-(3-(Aminomethyl)-4-methylphenyl)-3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea

Step 1

Using the procedure as described in Section 5.15,1-(3-cyano-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureais prepared from 5-amino-2-methylbenzonitrile and3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt.

Step 2

A mixture of1-(3-cyano-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea(0.54 g, 1.0 mmol), platinum oxide (0.1 g), and 5-6M HCl in isopropanol(2 mL) in acetic acid (15 mL) is hydrogenated under 50 psi hydrogen for48 hrs. The mixture is filtered through Celite, the filtrate isevaporated under vacuum, and the residue is purified by preparative HPLCto provide the product.

5.241-(2-(Aminomethyl)-5-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea

Step 1

Using the procedure as described in Section 5.15,1-(2-cyano-5-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureais prepared from 2-amino-4-methylbenzonitrile and3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt.

Step 2

A mixture of1-(2-cyano-5-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea(0.54 g, 1.0 mmol), platinum oxide (0.1 g), and 5-6M HCl in isopropanol(2 mL) in acetic acid (15 mL) is hydrogenated under 50 psi hydrogen for48 hrs. The mixture is filtered through Celite, the filtrate isevaporated under vacuum, and the residue is purified by preparative HPLCto provide the product.

5.251-((2(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-(morpholinomethyl)phenyl)urea

Using the procedure as described in Section 5.15, the product isprepared from 4-(morpholinomethyl)aniline and3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt.

5.261-((2-((2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-((4-methylpiperazin-1-yl)methyl)phenyl)urea

Using the procedure as described in Section 5.15, the product isprepared from 3-((4-methylpiperazin-1-yl)methyl)aniline and3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt.

5.27 Isoindolin Compounds

The isoindolin compounds shown above are made using the procedure asdescribed in Section 5.15.

5.281-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H′-isoindol-5-ylmethyl]-3-[4-(2-pyridin-4-yl-ethyl)-phenyl]-ureahydrochloric acid salt

Step 1: Preparation of 4-[2-(4-nitro-phenyl)-vinyl]-pyridine

To a CH₃CN solution of 1-iodo-4-nitrobenzene (498 mg, 2 mmol) and4-vinylpyridine (262 mg, 2.5 mmol) was added TEA (350 μL, 2.5 mmol) andPd(OAc), (0.45 mg, 0.2 mmol). The mixture was heated to 100° C. for 48hr in a capped Pyrex tube. It was then cooled to room temperature,quenched with 1N HCl (20 mL) and concentrated under reduced pressure.The mixture was filtered and the solid was taken up in EtOAc (30 mL) andwashed with 1N NaOH (30 mL). The organic layer was concentrated, driedover Na₂SO₄ and concentrated to give4-[2-(4-nitro-phenyl)-vinyl]-pyridine as yellow solid (160 mg, 37%).

Step 2: Preparation of 4-(2-pyridin-4-yl-ethyl)-phenylamine

To the EtOAc solution (30 mL) of 4-[2-(4-nitro-phenyl)-vinyl]-pyridine(160 mg, 0.71 mmol) was added palladium on carbon (0.1 g, 50% wet). Thesuspension was hydrogenated at 50 psi of hydrogen for 2 hours. Themixture was filtered over a celite pad. The filtrate was concentrate togive 4-(2-pyridin-4-yl-ethyl)-phenylamine as a yellow solid (140 mg,100%).

Step 3: Preparation of1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[4-(2-pyridin-4-yl-ethyl)-phenyl]-ureahydrochloric acid salt

To a suspension of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.37 g, 1 mmol) in DMF (2 mL) was addedcarbonyl diimidazole (162 mg, 1 mmol). The mixture was stirred at roomtemperature overnight. 4-Pyridin-4-ylethyl-phenylamine (140 mg, 0.7mmol) was added to the mixture and the mixture was stirred at 40° C.overnight then 80° C. for 1.5 hours. The mixture was cooled to roomtemperature, added water (5 mL) and stirred for 10 min. The suspensionwas filtered and the filtrate was concentrated and purified onprep-HPLC. The resulted solid after purification was stirred with 1NHCl, filtered and the filtrate was concentrated to give1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[4-(2-pyridin-4-yl-ethyl)-phenyl]-ureahydrochloric acid salt as an off-white solid (30 mg, 8% yield). HPLC:Waters Symmetry C₁₈, 4 μm, 3.9×150 mm, 1 mL/min, 240 nm, 5% grad 95% in5 min, CH₃CN/0.1% H₃PO₄: t_(R)=4.70 min (92%); mp: >400° C.; ¹H NMR(DMSO-d₆) δ 1.89-2.06 (m, 1H, CHH), 2.37 (br. s., 1H, CHH), 2.55-2.67(m, 1H, CHH), 2.82-3.01 (m, 3H, CH₂, CHH), 3.12 (d, J=7.7 Hz, 2H, CH₂),4.20-4.53 (m, 5H, CH₂, CH₂), 5.11 (dd, J=4.8, 13.3 Hz, 1H, NCH), 6.89(br. s., 1H, NH), 7.07 (d, J=8.3 Hz, 2H, Ar), 7.32 (d, J=8.3 Hz, 2H,Ar), 7.44 (d, J=7.9 Hz, 1H, Ar), 7.51 (s, 1H, Ar), 7.69 (d, J=7.7 Hz,1H, Ar), 7.83 (d, J=5.5 Hz, 2H, Ar), 8.71-8.78 (m, 2H, Ar), 8.80 (s, 1H,NH), 10.98 (s, 1H, NH). ¹³C NMR (DMSO-d₆) δ 22.44, 31.13, 34.25, 36.52,42.64, 47.05, 51.49, 117.59, 121.78, 122.84, 126.56, 126.78, 128.49,130.19, 132.39, 138.59, 142.12, 142.28, 144.86, 155.24, 167.86, 170.91,172.78; LCMS MH⁺=498; Anal Calcd for C₂₈H₂₇N₅O₄+1.55 HCl+0.75H₂O+0.15CH₃CN C, 56.78; H, 5.29; N, 11.64; Cl, 8.87. Found: C, 56.47; H, 5.15;N, 11.99; Cl, 8.85.

5.29 Isoindolin Compounds

These isoindolin compounds shown above are made using the procedure asdescribed in Section 5.15.

5.30N″-(3-Chloro-4-methyl-phenyl)-N′-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-N-cyano-guanidine

Step 1: Preparation of (3-chloro-4-methyl-phenyl)-carbamic acid phenylester

2-chloro-4-amino toluene (282 mg, 2 mmol) was dissolved in THF (10 mL).The mixture was added sodium hydride (128 mg, 3.2 mmol) and stirred atroom temperature for 15 minutes. Dipehnyl N-cyano-carbonimidate (715 mg,3.0 mmol) was added and the mixture was heated to reflux for 4 hours.The reaction mixture was cooled to room temperature, quenched bysaturated NH₄Cl (10 mL), filtered and the solid was dried in oven togive (3-chloro-4-methyl-phenyl)-carbamic acid phenyl ester as solid (0.5g, 87%).

Step 2: Preparation ofN″-(3-Chloro-4-methyl-phenyl)-N′-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-N-cyano-guanidine

To a suspension of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.18 g, 0.5 mmol) in DMF (2 mL), was added(3-chloro-4-methyl-phenyl)-carbamic acid phenyl ester (143 mg, 0.5 mmol)and DIPEA (83 μL, 0.5 mmol). The mixture was stirred at 100° C. for 4hours. The mixture was concentrated under reduced pressure to removeDMF, purified on silica gel column eluted using methanol and methylenechloride to giveN″-(3-Chloro-4-methyl-phenyl)-N′-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-N-cyano-guanidineas a white solid (40 mg, 17% yield). HPLC: Waters Symmetry C₁₈, 5 μm,3.9×150 mm, 1 mL/min, 240 nm, 50/50. CH₁CN/0.1% H₃PO₄: t_(R)=3.06 min(96.5%); mp: 325-327° C.; NMR (DMSO-d₆) δ 1.93-2.09 (m, 1H, CHH),2.25-2.32 (m, 3H, CH₃), 2.33-2.45 (m, 1H, CHH), 2.55-2.67 (m, 1H, CHH),2.82-3.01 (m, 1H, CHH), 4.27-4.57 (m, 4H, CH₂, CH₂), 5.11 (dd, J=5.1,13.2 Hz, 1H, NCH), 7.13 (dd, J=2.3, 8.1 Hz, 1H, Ar), 7.32 (d, J=1.5 Hz,2H, Ar), 7.43 (d, J=7.7 Hz, 1H, Ar), 7.51 (s, 1H, Ar), 7.71 (d, J=7.7Hz, 1H, Ar), 7.85 (s, 1H, NH), 9.20 (s, 1H, NH), 10.99 (s, 1H, NH). ¹³CNMR (DMSO-d₆) δ 18.98, 22.49, 31.20, 44.70, 47.15, 51.58, 116.88,121.98, 122.94, 124.33, 126.97, 130.52, 131.41, 132.08, 133.10, 136.46,142.36, 142.97, 158.15, 167.86, 170.98, 172.86.; LC-MS: 465; Anal Calcdfor C₂₃H₂₁ClN₆O₃+0.6 H₂O+0.2 EtOAc: C, 57.94; H, 4.86; N, 17.03. Found:C, 57.66; H, 4.81; N, 17.10.

5.311-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[4-(1H-imidazol-2-yl)-phenyl]-urea

A mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.40 g, 1.1 mmol) and CDI (0.19 g, 1.2 mmol)in DMF (5 mL) was stirred at room temperature for 18 hours. To themixture was added 4-(1′-1-imidazol-2-yl)-phenylamine (0.17 g, 1.1 mmol)at room temperature, and the mixture was stirred at 60° C. for 1d. Tothe mixture was added water (25 mL) and ethyl acetate (20 mL). Themixture was stirred at room temperature for 2 h. The suspension wasfiltered and the solid was washed with water (20 mL), ethyl acetate (20mL) and water (20 mL) to give a solid. The solid was purified with PrepHPLC to give1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[4-(1H-imidazol-2-yl)-phenyl]-ureaas a white solid (150 mg, 30% yield): HPLC: Waters Symmetry C₁₈, 5 μm,3.9×150 mm, 1 mL/min, 240 nm, 5/95 grad 95/5 in 5 min CH₁CN/0.1% H₃PO₄,4.41 min (95.9%); mp: 190-192° C.; ¹H NMR (DMSO-d₆) δ 1.91-2.08 (m, 1H,CHH), 2.27-2.47 (m, 1H, CHH), 2.54-2.66 (m, 1H, CHH), 2.81-3.01 (m, 1HCHH), 4.31 (d, J=17.4 Hz, 1H, CHH), 4.38-4.52 (m, 3H, CHH, CH₂), 5.11(dd, J=5.0, 13.3 Hz, 1H, NCH), 6.78 (t, J=6.0 Hz, 1H, NH), 7.33-7.76 (m,9H, Ar), 8.17 (s, 1H, HCOOH), 8.67 (s, 1H, NH), 10.99 (s, 1H, NH); ¹³CNMR (DMSO-d₆) δ 22.41, 31.10, 42.69, 47.02, 51.49, 114.17 (br), 117.80,121.79, 122.84, 124.55, 126.81, 126.98, 130.19, 135.46, 138.55, 142.31,144.83, 155.14, 163.19 (HCOOH), 167.88, 170.92, 172.77; LCMS MH=459:Anal. Calcd for C₂₄H₂₂N₆O₄+2 H₂O+0.7 HCOOH+0.3 DMF: C, 56.04; H, 5.42;N, 16.08. Found: C, 55.84; H, 5.34; N, 16.11.

5.321-[3-(1H-Benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea

A mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.40 g, 1.1 mmol) and CDI (0.21 g, 1.3 mmol)in DMF (5 mL) was stirred at room temperature for 18 h. To the mixturewas added 3-(1H-benzoimidazol-2-yl)-4-chloro-phenylamine (0.32 g, 1.3mmol) at room temperature, and the mixture was stirred at 100° C. for 18hours. To the mixture was added water (25 mL) and ether (20 mL). Themixture was stirred at room temperature for 2 h. The solvent wasdecanted. The solid was purified with Prep HPLC to give1-[3-(1H-benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureaas a white solid (130 mg, 22% yield): HPLC: Waters Symmetry C₁₈, 5 μm,3.9×150 mm, 1 mL/min, 240 nm, 20/80 CH₃CN/0.1% H₃PO₄, 4.44 min (99.2%);mp: 275-277° C.; ¹H NMR (DMSO-d₆) δ 1.90-2.10 (m, 1H, CHH), 2.23-2.46(m, 1H, CHH), 2.54-2.68 (m, 1H, CHH), 2.80-3.02 (m, 1H, CHH), 4.31 (d,J=17.4 Hz, 1H, CHH), 4.37-4.51 (m, 3H, CHH, CH₂), 5.11 (dd, J=5.0, 13.3Hz, 1H, NCH), 6.89 (t, J=5.9 Hz, 1H, NH), 7.23 (d, J=4.5 Hz, 2H, Ar),7.40-7.76 (m, 7H, Ar), 8.07 (d, J=2.5 Hz, 1H, Ar), 9.02 (s, 1H, NH),10.98 (s, 1H, NH), 12.64 (br. s., 1H, NH); ¹³C NMR (DMSO-d₆) δ 22.39,31.10, 42.75, 47.02, 51.48, 111.62, 118.88, 120.11, 120.62, 121.55,121.83, 122.56, 122.85, 126.85, 129.79, 130.23, 130.35, 134.56, 139.56,142.29, 143.00, 144.58, 149.12, 154.96, 162.96, 167.86, 170.91, 172.76;LCMS MH=543, 545; Anal. Calcd for C₂₈H₂₃N₆O₄Cl+3H₂O: C, 56.33; H, 4.90;N, 14.08. Found: C, 56.73; H, 4.67; N, 14.09.

5.33N-(3-Chloro-4-methyl-phenyl)-N′-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-guanidineHydrochloride

Step 1: 3-Chloro-4-methyl-phenyl)-thiourea

To a solution of 2-chloro-4-isothiocyanato-1-methyl-benzene (2 gm, 10.89mmol) in dry acetonitrile (10 mL) was added a solution of NH₃ in MeOH (2N, 6 mL) and the mixture was stirred at rt for 2 h. Water was added tothe reaction mixture causing the a precipitate to form. The reactionslurry was concentrated in vacuo to ˜1/4 the volume and the white solidwas collected by filtration and dried in a vacuum oven to give 2.1 gm(96% yield) of 3-chloro-4-methyl-phenyl)-thiourea as a white solid thatwas used without further purification. LCMS: MH=201.

Step 2: 1-(3-Chloro-4-methyl-phenyl)-2-methyl-isothiourea Hydroiodide

To a slurry of 3-chloro-4-methyl-phenyl)-thiourea (2.1 gm, 10.46 mmol)in MeOH (15 mL) was added methyl iodide (2.5 mL, 40 mmol) and themixture was stirred at rt for 4 h. The reaction mixture was concentratedto dryness and to the residue, a small portion of dichloromethane wasadded and the volatiles removed in vacuo. This process was repeatedtwice more to provide a tan foam which was dried in a vacuum ovenovernight to give 3.5 gm (98% yield) of crude (2×).1-(3-chloro-4-methyl-phenyl)-2-methyl-isothiourea hydroiodide. Thismaterial was found to be highly hygroscopic and was stored in adesiccator for further use. LCMS: MH=215.

Step 3:N-(3-Chloro-4-methyl-phenyl)-N′-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-guanidineHydrochloride

In a microwave vial fitted with a stir bar,1-(3-chloro-4-methyl-phenyl)-2-methyl-isothiourea hydroiodide (597 mg,1.74 mmol),3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonate (560 mg, 1.52 mmol), and DIEA (1.1 mL, 6.06 mmol) werecombined in anhydrous DMF (5 mL). The vial was sealed and irradiated ina microwave for 30 min at 120° C. To the reaction mixture was added anadditional 100 mg of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonate and the reaction was irradiated for 30 min at 120° C.This process was repeated twice more to consume all of the1-(3-chloro-4-methyl-phenyl)-2-methyl-isothiourea hydroiodide startingmaterial as it was found co-elute with the desired product on LCMS. Thereaction mixture was acidified with acetic acid and the volatiles wereevaporated in vacuo. The residue was dissolved in minimal DMF, filtered,and purified using C-18 preparatory HPLC. To the combined fractionscontaining the desired product, 1 N HCl was added and the solventsremoved in vacuo. The obtained white solid was dissolved in minimalwater and lyophilized to give 180 mg (27% yield) ofN-(3-chloro-4-methyl-phenyl)-N′-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-guanidinehydrochloride: HPLC:Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 ml/min, 240nm, 20/80 CH₃CN/0.1% H₃PO₄, 3.73 min (97.4%); mp: >400° C.; ¹H NMR(DMSO-d₆) 1.91-2.09 (m, 1H, CHH), 2.33 (s, 3H, CH₃), 2.41 (dd, J=4.3,13.2 Hz, 1H, CHH), 2.54-2.68 (m, 1H, CHH), 2.80-3.05 (m, 1H, CHH),4.21-4.56 (m, 2H, CH₂), 4.66 (d, J=5.7 Hz, 2H, CH₂), 5.13 (dd, J=4.9,13.2 Hz, 1H, CH), 7.14 (dd, J=1.9, 8.1 Hz, 1H, Ar), 7.34 (d, J=1.9 Hz,1H, Ar), 7.42 (d, J=8.1 Hz, 1H, Ar), 7.51 (d, J=7.9 Hz, 1H, Ar), 7.61(s, 1H, Ar), 7.76 (d, J=7.7 Hz, 1H, Ar), 7.98 (br. s., 1H, NH), 8.62(br. s., 1H, NH), 10.16 (s, 1H, NH), 11.00 (s, 1H, NH); ¹³C NMR(DMSO-d₆) 21.04, 24.19, 32.95, 46.33, 49.85, 54.03, 124.13, 125.48,125.79, 127.28, 129.17, 132.57, 134.20, 135.82, 135.95, 136.64, 143.17,144.68, 156.84, 170.94, 173.00, 175.82; LCMS: MH=440, 442; Anal Calcdfor C₂₂H₂₃Cl₂N₅O₃+2.3 H₂O+1.9 HCl+0.2 HCOOH: C, 44.72; H, 5.05; N,11.74; Cl, 23.19. Found: C, 44.55; H, 4.71; N, 11.48; Cl, 22.92.

5.341-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-ylmethyl)-3-(3-hydroxy-4-methylphenyl)urea

Step 1

A mixture of 2-methyl-5-nitrophenol (3.0 g, 19.6 mmol) TBS-Cl (3.0 g,19.6 mmol), and TEA (2.2 g, 21.6 mmol) in DMF (30 mL) stirred at ambienttemperature for 16 h. The mixture was evaporated under vacuum. Theresidue was dissolved in ethyl acetate (100 mL) and washed with water(3×100 mL) and evaporated under vacuum. The residue was chromatographedin a hexanes-ethyl acetate gradient, providing 4.7 g oftert-butyldimethyl(2-methyl-5-nitrophenoxy)silane, in 91% yield; ¹H NMR(DMSO-d₆) δ 0.27 (s, 6H), 1.01 (s, 9H), 2.27 (s, 3H), 7.47 (d, J=8.5 Hz,1H), 7.52 (d, J=2.3 Hz, 1H) 7.78 (dd, J=2.3, 8.3 Hz, 1H).

Step 2

A mixture of the product from Step 1 (4.7 g, 17.6 mmol) and 10% Pd—C(1.0 g, 50% wet) in ethyl acetate was hydrogenated under 50 psi hydrogenfor 16 h. The mixture was then filtered through Celite and the filtratewas evaporated under vacuum, providing 3.9 g of-(tert-butyldimethylsilyloxy)-4-methylaniline, in 94% yield; ¹H NMR(DMSO-d₆) δ 0.17 (s, 6H), 0.97 (s, 9H), 1.96 (s, 3H), 4.81 (s, 2H),6.00-6.21 (m, 2H), 6.74 (d, J=7.9 Hz, 1H).

Step 3

A mixture of3-(5-(aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dionemethanesulfonate (0.74 g, 2.0 mmol) and CDI (0.32 g, 2.0 mmol) in DMF(30 mL) was stirred at room temperature for 3 h, and then the productfrom Step 2 (0.47 g, 2.0 mmol) was added. The mixture was heated to 70°C. for 16 h. Then, the mixture was cooled to room temperature andquenched with 1N HCl (30 mL). The resulting mixture was evaporated undervacuum and the residue was purified by preparative HPLC using anacetonitrile-water gradient, and providing 0.38 g of1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)ureaas a white solid, in 46% yield; HPLC:Waters Symmetry C₁₈, 5 μm, 3.9×150mm, 1 ml/min, 240 nm, 25/75 CH₃CN/0.1% H₃PO₄, 4.80 min (100.00%); mp273-275° C.; ¹H NMR (DMSO-d₆) δ 2.02 (s, 4H), 2.27-2.47 (m, 1H), 2.63(m., 1H), 2.80-3.04 (m, 1H), 4.17-4.59 (m, 4H), 5.11 (dd, J=4.7, 13.0Hz, 1H), 6.47-6.72 (m, 2H), 6.86 (d, J=7.9 Hz, 1H), 7.04 (s, 1H), 7.44(d, J=7.7 Hz, 1H), 7.51 (s, 1H), 7.69 (d, J=7.7 Hz, 1H), 8.39 (s, 1H),9.13 (s, 1H), 10.98 (s, 1H); ¹³C NMR (DMSO-d₆) δ 15.35, 22.49, 31.20,42.74, 47.12, 51.58, 104.76, 108.45, 116.48, 121.86, 122.91, 126.91,130.19, 130.26, 138.92, 142.36, 144.96, 155.18, 155.29, 167.95, 170.99,172.85; LCMS MH=423; Anal. Calcd for C₂₂H₂₂N₄O₅: C, 62.55; H, 5.25; N,13.26. Found: C, 62.27; H, 5.15; N, 13.18.

5.351-((2-(2,6-Dioxopiperidin-3-O-1-oxoisoindolin-5-yl)methyl)-3-(4-hydroxy-3-methylphenyl)urea

Step 1

A mixture of 2-methyl-4-nitrophenol (3.0 g, 19.6 mmol), TBS-Cl (3.0 g,19.6 mmol), and TEA (2.2 g, 21.6 mmol) in DMF (30 mL) stirred at ambienttemperature for 16 h. The mixture was partitioned between 10% aqueoussodium bicarbonate solution (100 mL) and ethyl acetate (100 mL), and theaqueous layer was extracted with ethyl acetate (100 mL). The combinedorganic layers were washed with 10% aqueous sodium bicarbonate (3×100mL) and water (100 mL), and evaporated under vacuum. The residue waschromatographed in hexanes-ethyl acetate gradient, providing 3.9 g oftert-butyldimethyl(2-methyl-4-nitrophenoxy)silane, in 75% yield; ¹H NMR(DMSO-d₆) δ 0.29 (s, 6H), 1.00 (s, 9H), 2.24 (s, 3H), 7.03 (d, J=8.9 Hz,1H), 8.02 (dd, J=2.8, 8.9 Hz, 1H), 8.12 (d. J=2.5 Hz, 1H).

Step 2

A mixture of the product from step 1 (3.9 g, 14.6 mmol) and 10% Pd—C(0.5 g, 50% wet) in ethyl acetate was hydrogenated under 50 psi hydrogenfor 18 h. The mixture was filtered through Celite and the filtrate wasevaporated, providing 3.2 g of4-(tert-butyldimethylsilyloxy)-3-methylaniline, in 93% yield.

Step 3

A mixture of3-(5-(aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dionemethanesulfonate (0.74 g, 2.0 mmol) and CDI (0.32 g, 2.0 mmol) in DMF(30 mL) was stirred at room temperature for 3 h, and then the productfrom Step 2 (0.47 g, 2.0 mmol) was added. The mixture was heated to 70°C. for 30 h. Then, the mixture was cooled to room temperature andquenched with 1N HCl (30 mL), resulting in solid precipitate. This solidwas purified by chromotagraphy on a silica gel column, using a methylenechloride-methanol gradient, and providing 0.59 g of1-(4-(tert-butyldimethylsilyloxy)-3-methylphenyl)-3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureaas an off-white solid, in 55% yield; ¹H NMR (DMSO-d₆) δ 0.16 (s, 6H),0.97 (s, 9H), 1.88-2.06 (m, 1H), 2.10 (s, 3H), 2.25-2.47 (m, 1H),2.53-2.70 (m, 1H), 2.80-3.02 (m, 1H), 4.31 (d, J=17.4 Hz, 1H), 4.36-4.52(m, 3H), 5.11 (dd, J=4.9, 13.2 Hz, 1H), 6.51-6.74 (m, 2H), 7.08 (dd,J=2.5, 8.6 Hz, 1H), 7.18 (d, J=2.3 Hz, 1H), 7.43 (d, J=7.7 Hz, 1H), 7.50(s, 1H), 7.69 (d, J=7.7 Hz, 1H), 8.35 (s, 1H), 10.98 (s, 1H).

Step 4

To a solution of the product from Step 3 (0.5 g, 0.9 mmol) in methylenechloride (20 mL) was added 2N HCl in ether (2 mL). The mixture stirredfor 16 h at ambient temperature. The precipitated product was isolatedby filtration, and was rinsed with methylene chloride (20 mL) and driedunder vacuum, providing 0.4 g of1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-hydroxy-3-methylphenyl)ureaas an off-white solid, in quantitative yield; HPLC:Waters Symmetry C₁₈,5 μm, 3.9×150 mm, 1 ml/min, 240 nm, 30/70 CH₃CN/0.1% H₃PO₄, 1.88 min(96.94%); mp 210-212° C.; ¹H NMR (DMSO-d₆) δ 1.94-2.18 (m, 4H),2.30-2.51 (m, 1H), 2.58-2.72 (m, 1H), 2.87-3.06 (m, 1H), 4.25-4.56 (m,4H), 5.15 (dd, J=5.0, 13.3 Hz, 1H), 6.67 (d, J=8.5 Hz, 1H), 7.02 (dd,J=2.5, 8.6 Hz, 1H), 7.12 (d, J=2.3 Hz, 1H), 7.47 (d, J=7.7 Hz, 1H), 7.54(s, 1H), 7.73 (d, J=7.7 Hz, 1H), 8.32 (br. s., 1H), 11.02 (s, 1H); ¹³CNMR (DMSO-d₆) δ 16.15, 22.51, 31.20, 42.77, 47.12, 51.56, 114.47,117.11, 121.32, 121.83, 122.88, 123.61, 126.86, 130.22, 131.72, 142.35,145.17, 150.10, 155.58, 170.99, 172.85; LCMS MH=423; Anal. Calcd forC₂₂H₂₂N₄O₅+0.25 CH₂Cl₂: C, 60.23; H, 5.11; N, 12.63. Found: C, 60.14; H,5.43; N, 12.54.

5.361-(4-tert-Butyl-cyclohexyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-4H-isoindol-5-ylmethyl]-urea

To a stirred suspension of 4-tert-butyl-cyclohexylamine (0.20 g, 1.28mmol) in DMF (5 mL) at 40° C. was added CDI (0.23 g, 1.40 mmol). Themixture was stirred for 15 min, followed by addition of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.47 g, 1.28 mmol). Heating was stopped after1.5 hrs and the mixture was stirred at RT overnight. Solvent wasevaporated and the residue was purified by preparative HPLC to give theproduct as a white solid (0.076 g, 13% yield):

HPLC, Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 ml/min, 240 nm, 35/65CH₃CN/0.1% H₃PO₄, 13.37 min (44.7%), 14.68 min (54.3%); mp, 182-184° C.;¹H NMR (DMSO-d₆) δ 0.83 (s, 9H, CH₃, CH₃, CH₃), 0.90-1.21 (m, 4H, CH₂,CH₂), 1.30-1.58 (m, 2H, CHH, CHH), 1.61-1.78 (m, 2H, CHH, CHH), 1.87(br. s., 1H, CHH), 1.95-2.06 (m, 1H, CHH), 2.28-2.47 (m, 1H, CHH), 2.63(br. s., 1H, CHH), 2.83-3.00 (m, 1H, CHH), 3.78 & 3.27 (2s, 1H, CH),4.23-4.50 (m, 4H, CH₂, CH₂), 5.11 (dd, J=5.0, 13.1 Hz, 1H, NCH),5.79-6.12 (m, 1H, NH), 6.25-6.44 (m, 1H, NH), 7.38 (t, 6.8 Hz, 1H, ArH),7.45 (d, J=6.2 Hz, 1H, ArH), 7.67 (dd, J=4.1, 7.6 Hz, 1H, ArH), 10.99(s, 1H, NH); (Note: ¹H NMR showed about 55% to 45% isomer ratio); ¹³CNMR (DMSO-d₆) δ 21.33, 22.49, 25.95, 27.38, 27.45, 31.02, 31.20, 32.06,32.26, 33.76, 42.87, 43.43, 46.81, 47.10, 47.42, 48.69, 51.56, 121.76,121.88, 122.83, 122.90, 126.81, 126.88, 130.11, 130.19, 142.29, 142.35,145.36, 145.52, 157.35, 167.98, 170.99, 172.85. LC/MS MH⁺=455; Anal.Calcd. For C₂₅H₃₄N₄O₄+0.7 H₂O: C, 64.27; H, 7.64; N, 11.99. Found: C,63.98; H, 7.98; N, 11.92.

5.371-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea

To a stirred suspension of 4-methyl-cyclohexylamine (mixture ofcis/trans isomers, 0.21 g, 1.86 mmol) in DMF (10 mL) at 40° C. was addedCDI (0.33 g, 2.04 mmol). The mixture was stirred for 15 min, followed byaddition of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.69 g, 1.86 mmol). Heating was stopped after1.5 hrs and the mixture was stirred at RT overnight. Solvent wasevaporated and the residue was purified by preparative HPLC to give theproduct as a white solid (0.14 g, 18% yield): HPLC, Waters Symmetry C₁₈,5 μm, 3.9×150 mm, 1 ml/min, 240 nm, 25/75 CH₃CN/0.1% H₃PO₄, 11.33 min(59.1%), 12.41 min (39.7%); mp, 223-225° C.; ¹H NMR (DMSO-d₆) δ0.81-0.93 (m, 3H, CH₃), 0.95-1.35 (m, 3H, CHH, CHH, CHH), 1.37-1.57 (m,4H, CHH, CHH, CHH, CH), 1.58-1.70 (m, 1H, CHH), 1.73-1.88 (m, 1H, CHH),1.92-2.07 (m, 1H, CHH), 2.30-2.47 (m, 1H, CHH), 2.54-2.67 (m, 1H, CHH),2.83-3.00 (m, 1H, CHH), 3.20-3.75 (m, 1H, CH), 4.21-4.51 (m, 4H, CH₂,CH₂), 5.11 (dd, J=5.0, 13.1 Hz, 1H, NCH), 5.78-6.11 (m, 1H, NH),6.24-6.40 (m, 1H, NH), 7.34-7.42 (m, 1H. ArH), 7.42-7.50 (m, 1H, ArH),7.63-7.72 (m, 1H, ArH), 10.98 (s, 1H, NH); (Note: ¹H NMR showed about60% to 40% isomer ratio); ¹³C NMR (DMSO-d₆) δ 21.47, 22.17, 22.49,29.42, 29.77, 30.42, 31.20, 31.50, 33.28, 33.73, 42.86, 44.54, 47.09,48.37, 51.56, 121.78, 122.87, 126.81, 130.16, 142.33, 145.44, 145.52,157.33, 167.98, 170.99, 172.85; LC/MS MH⁺=413; Anal. Calcd. ForC₂₂H₂₈N₄O₄+0.4 H₂O: C, 63.23; H, 6.90; N, 13.41. Found: C, 62.93; H,6.92; N, 13.09.

5.381-(3-Diethylamino-propyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea,formic acid salt

To the solution of para-nitrophenyl chloroformate (202 mg, 1 mmol) inCH₃CN (5 mL) was added dropwise at 0° C. a solution ofN,N-diethyl-propane-1,3-diamine (130 mg, 1 mmol) and DIPEA (0.082 mL, 1mmol) in CH₃CN (5 mL). The mixture was stirred at 0° C. for 10 min.345-Aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.37 g, 1 mmol) was added, followed byadditional DIPEA (0.082 mL, 1 mmol). The mixture was allowed to warm toRT and stirred overnight at RT. The mixture was added 1N HCl (15 mL) andfiltered. The resulted solid was purified on preparative HPLC to givethe product as an off-white solid (280 mg, 65% yield): HPLC: WatersSymmetry C₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm, 5% gradient 100% in 5min, CF₃CN/0.1% H₃PO₄, 4.46 min (97%); mp: 90-92° C.; ¹H NMR (DMSO-d₆) δ1.01 (t, J=7.1 Hz, 6H, CH₃, CH₃), 1.58 (quin, J=7.1 Hz, 2H, CH₂),1.94-2.06 (m, 1H, CHH), 2.39 (qd, J=4.5, 13.2 Hz, 1H, CHH), 2.55-2.73(m, 7H, CH₂, CH₂, CH₂, CHH), 2.83-2.99 (m, 1H, CHH), 3.05 (d, J=5.3 Hz,2H, CH₂), 4.15-4.56 (m, 4H, CH₂, CH₂), 5.10 (dd, J=5.1, 13.4 Hz, 1H,NCH), 6.17 (br. s., 1H, NH), 6.60 (s, 1H, NH), 7.39 (d, J=7.7 Hz, 1H,Ar), 7.45 (s, 1H, Ar), 7.67 (d, J=7.9 Hz, 1H, Ar), 8.25 (br. s., 1H,HCOOH), 10.98 (br. s., 1H, NH); ¹³C NMR (DMSO-d₆) δ 10.25, 22.49, 26.14,31.20, 37.39, 42.93, 46.11, 47.09, 51.56, 121.73, 122.81, 126.76,130.13, 142.27, 145.54, 158.18, 167.99, 171.01, 172.86; LCMS MH⁺=430;Anal. Calcd. for C₂₃H₃₁N₅O₆+2 H₂O: C, 54.00; H, 7.29; N, 13.69. Found:C, 54.04; H, 6.92; N, 13.63.

5.391-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-methyl-tetrahydro-pyran-4-yl)-urea

To the solution of para-nitrophenyl chloroformate (202 mg, 1 mmol) inCH₃CN (5 mL) was added dropwise at 0° C. a solution of4-methyl-tetrahydro-pyran-4-ylamine (150 mg, 1 mmol) and DIPEA (0.082mL, 1 mmol) in CH₃CN (5 mL). The mixture was stirred at 0° C. for 10min.3-(5-Aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.37 g, 1 mmol) was added, followed byadditional DIPEA (0.082 mL, 1 mmol). The mixture was allowed to warm toRT and stirred overnight at RT. The mixture was added 1N HCl (15 mL) andfiltered. The resulted solid was purified on preparative HPLC to give asolid. The solid was recrystallized from DMF (1 mL) and water (1 mL) togive the product as a white solid (55 mg, 12% yield): HPLC: WatersSymmetry C₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm, 5% gradient 100% in 5min. CH₃CN/0.1% H₃PO₄, 5.02 min (98%); mp: 158-160° C.; ¹H NMR (DMSO-d₆)δ 1.29 (s, 3H, CH₃), 1.40-1.58 (m, 2H, CHH), 1.84-1.96 (m, 2H, CHH,CHH), 1.99 (s, 1H, CHH), 2.30-2.45 (m, 1H, CHH), 2.55-2.65 (m, 1H, CHH),2.84-2.99 (m, 1H, CHH), 3.44-3.68 (m, 4H, CH₂, CH₂), 4.22-4.58 (m, 4H,CH₂, CH₂), 4.95-5.29 (m, 1H, NCH), 5.85 (s, 1H, NH), 6.08-6.52 (m, 1H,NH), 7.33-7.42 (m, 1H. Ar), 7.45 (s, 1H, Ar), 7.68 (d, J=7.6 Hz, 1H.Ar), 10.98 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 22.48, 27.00, 31.20, 36.93,42.58, 47.10, 48.85, 51.56, 62.92, 120.99, 121.67, 122.88, 126.69,142.35, 145.48, 157.22, 167.98, 171.01, 172.86; LCMS MH⁺=415; Anal.Calcd. for C₂₁H₂₆N₄O₅+0.5 H₂O: C, 59.56; H, 6.43; N, 13.23. Found: C,59.34; H, 6.35; N, 13.21.

5.401-(1-Benzyl-piperidin-4-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea

To the solution of para-nitrophenyl chloroformate (202 mg, 1 mmol) inCH₃CN (5 mL) was added dropwise at 0° C. a solution of4-amino-benzyl-piperidine (190 mg, 1 mmol) and DIPEA (0.082 mL, 1 mmol)in CH₃CN (5 mL). The mixture was stirred at 0° C. for 10 min.3-(5-Aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.37 g, 1 mmol) was added, followed byadditional DIPEA (0.082 mL, 1 mmol). The mixture was allowed to warm toRT and stirred overnight at RT. The mixture was added 1N HCl (15 mL) andfiltered. The resulted solid was purified on preparative-HPLC to give agluey solid. The solid was dissolved in DMF (1 mL) and saturated NaHCO₃solution was added dropwise until precipitation started occurring. Itwas sat still at RT for 30 min and solid was filtered to give theproduct as a white solid (55 mg, 12% yield): HPLC: Waters Symmetry C₁₈,5 μm, 3.9×150 mm, 1 mL/min, 240 nm, 5% gradient 100% in 5 min,CH₃CN/0.1% H₃PO₄, 4.64 min (99%); mp: 208-210° C.; ¹H NMR (DMSO-d₆) δ1.21-1.52 (m, 2H, CHH), 1.63-1.82 (m, 2H, CHH), 1.89-2.14 (m, 3H, CHH,CHH, CHH), 2.39 (d, J=13.6 Hz, 1H, CHH), 2.55-2.65 (m, 1H, CHH),2.64-2.75 (m, 2H, CHH, CHH), 2.83-3.02 (m, 1H, CHH), 3.37-3.41 (m, 1H,CH), 3.43 (s, 2H, CH₂), 4.21-4.50 (m, 3H, CH₂, CH₂), 5.10 (dd, J=5.0,13.3 Hz, 1H, NCH), 5.94 (d, J=7.9 Hz, 1H, NH), 6.34 (t. J=6.0 Hz, 1H,NH), 7.13-7.48 (m, 6H, Ar), 7.66 (d, J=7.6 Hz, 1H, Ar), 10.97 (s, 1H,NH); ¹³C NMR (DMSO-d₆) δ 22.49, 31.20, 32.46, 42.86, 46.43, 47.09,51.55, 51.90, 62.19, 121.78, 122.84, 126.79, 128.10, 128.69, 130.13,138.64, 142.29, 145.44, 157.32, 167.96, 170.99, 172.85; LCMS MH⁺=490;Anal. Calcd. for C₂₁H₂₆N₄O₅+0.5 H₂O: C, 59.56; H, 6.43; N, 13.23. Found:C, 59.34; H, 6.35; N, 13.21.

5.411-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-piperidin-4-yl-ureahydrochloride salt

Step 1

A mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (1.25 g, 3.39 mmol) and CDI (0.61 g, 3.73mmol) in DMF (10 mL) was stirred at RT overnight.4-Amino-piperidine-1-carboxylic acid tert-butyl ester (0.68 g, 3.39mmol) was then added and the mixture was stirred overnight. The mixturewas purified by preparative HPLC to give4-{3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-piperidine-1-carboxylicacid tert-butyl ester as an off-white solid (0.46 g, 27% yield). Theproduct was used in the next step without further purification. HPLC,Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 ml/min, 240 nm, 30/70CH₃CN/0.1% H₃PO₄, 4.35 min (99.4%).

Step 2

A mixture of4-{3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-piperidine-1-carboxylicacid tert-butyl ester (0.45 g, 0.90 mmol) in 2 M hydrochloride indiethyl ether (30 mL, 9.01 mmol) was stirred overnight. The whitesuspension was filtered and washed with diethyl ether. The resultingwhite solid was stirred in acetonitrile (80 mL) at 50° C. overnight. Thesuspension was filtered, washed with acetonitrile, and vacuum dried togive the product as a white solid (0.34 g, 86% yield): HPLC:WatersXterra C₁₈, 5 μm, 3.9×150 mm, 1 ml/min, 240 nm, 5/95 CH₃CN/0.1% HCOONH₄gradient to 95/5 in 5 min, kept 5 min, 3.10 min (95.1%); mp, 340-342°C.; ¹H NMR (DMSO-d₆) δ 1.46-1.65 (m, 2H, CHH, CHH), 1.85-2.05 (m, 3H,CHH, CHH, CHH), 2.30-2.47 (m, 1H, CHH), 2.55-2.67 (m, 1H, CHH),2.83-3.03 (m, 3H, CHH, CHH, CHH), 3.15-3.27 (m, 2H, CHH, CHH), 3.59-3.77(m, 1H, CH), 4.23-4.51 (m, 4H, CH₂, CH₂), 5.11 (dd, J=4.9, 13.2 Hz, 1H,NCH), 6.40-6.57 (m, 2H, NH, NH), 7.39 (d, J=7.9 Hz, 1H, ArH), 7.45 (s,1H, ArH), 7.67 (d, J=7.7 Hz, 1H, ArH), 8.75 (br. s., 2H, ClH₂N), 10.98(s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 22.49, 28.92, 31.20, 41.96, 42.82,43.95, 47.09, 51.55, 121.76, 122.84, 126.76, 130.14, 142.29, 145.35,157.32, 167.98, 171.01, 172.87; LC/MS MH⁺=400; Anal. Calcd. ForC₂₀H₂₆N₅O₄Cl+1.7 H₂O: C, 51.49; H, 6.35; N, 15.01; Cl, 7.60. Found: C,51.18; H, 6.15; N, 14.90; Cl, 7.51.

5.42 Isoindolin Compounds

The isoindolin compounds shown above are using the procedure asdescribed in Section 5.15.

5.434-{3-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-cyclohexanecarboxylicacid amide

To the solution of para-nitro-phenyl chloroformate (200 mg, 1 mmol) inCH₃CN (5 mL), was added the CH₃CN solution (5 mL) of4-Amino-cyclohexanecarboxylic acid amide (210 mg, 1 mmol) and DIPEA (0.3mL, 2 mmol) dropwise at room temperature. The mixture was stirred atroom temperature for 10 min.3-(5-Aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.37 g, 1 mmol) was added followed byadditional DIPEA (0.15 mL, 1 mmol). The mixture was stirred overnight atambient temperature. The mixture was then filtered. The filtrate waspurified on prep-HPLC to give4-{3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-cyclohexanecarboxylicacid amide as a white solid (130 mg, 30% yield). HPLC: Waters SymmetryC₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm, 5% grad 95% in 5 min,CH₁CN/0.1% H₃PO₄: t_(R)=4.78 min (92%); mp: 195-197° C.; ¹H NMR(DMSO-d₆) 1.32-1.73 (m, 8H, CHH+CHH+CHH+CHH), 1.97 (br. S., 1H, CHH),2.09-2.21 (m, 1H, CHH), 2.40 (d, J=13.4 Hz, 1H, CHH), 2.62 (br. S., 1H,CHH), 2.80-3.03 (m, 1H, CHH), 3.70 (br. S., 1H, CHH), 4.18-4.58 (m, 4H,CH, +CH₂), 5.11 (dd, 1H, CHN), 6.15 (d, J=7.9 Hz, 1H, NH), 6.38 (t,J=5.9 Hz, 1H, NH), 6.67 (br. S., 1H, NH), 7.19 (br. S, 1H, NH), 7.38 (d,J=7.7 Hz, 1H, Ar), 7.45 (s, 1H, Ar), 7.67 (d, J=7.9 Hz, 1H, Ar), 10.98(br. S., 1H, NH); ¹³C NMR (DMSO-d₆) δ 0.03, 22.42, 24.06, 24.19, 29.73,41.73, 42.79, 44.16, 47.03, 51.49, 121.68, 122.80, 126.71, 130.09,142.26, 145.37, 157.28, 163.37, 167.89, 170.92, 172.78, 176.83; LC-MS:442; Anal Calcd for C₂₂H₂₇N₅O₅+0.5 HCOOH+1H₂O+0.2 CH₃CN: C, 56.05; H,6.21; N, 14.84. Found: C, 55.70; H, 6.46; N, 14.78.

5.441-(6-Chloro-pyridin-3-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea

To a suspension of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.37 g, 1 mmol) in DMF (3 mL) was addedcarbonyl diimidazole (162 mg, 1 mmol). The mixture was stirred at roomtemperature overnight. 5-Amino-2-chloro-pyridine (128.5 mg, 1 mmol) wasadded to the mixture and the mixture was heated at 80° C. for 6 hours.The mixture was then concentrated under reduced pressure and theresulted mixture was purified on silica gel column eluted with methylenechloride and methanol to give11-(6-chloro-pyridin-3-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureaas an off-white solid (20 mg, 5% yield): HPLC: Waters Symmetry C₁₈, 5μm, 3.9×150 mm, 1 mL/min, 240 nm, 5% grad 95% in 5 min, CH₃CN/0.1%H₃PO₄, t_(R)=5.43 min (97%); mp: 224-226° C.; ¹H NMR (DMSO-d₆) δ1.89-2.09 (m, 1H, CHH), 2.29-2.44 (m, 1H, CHH), 2.54-2.66 (m, 1H, CHH),2.83-3.04 (m, 1H, CHH), 4.20-4.57 (m, 4H, CH₂, CH₂), 5.11 (dd, J=5.0,13.3 Hz, 1H, NCH), 6.99 (t. J=6.2 Hz, 1H, NH), 7.37 (d, J=8.5 Hz, 1H,Ar), 7.45 (d, J=7.7 Hz, 1H, Ar), 7.52 (s, 1H, Ar), 7.70 (d, J=7.7 Hz,1H, Ar), 7.95 (dd, J=2.8, 8.7 Hz, 1H, Ar), 8.42 (d, J=2.3 Hz, 1H, Ar),9.03 (s, 1H, NH), 10.98 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 22.49, 31.18,42.86, 47.12, 51.58, 121.91, 122.94, 123.89, 126.92, 128.37, 130.42,136.75, 139.04, 142.39, 144.50, 154.97, 167.93, 171.01, 172.86, 215.61;LC-MS: 428; Anal Calcd for C₂₀H₁₈N₅O₄Cl+0.1 H₂O C, 55.91; H, 4.27; N,16.30; Cl, 8.25. Found: C, 56.31; H, 4.51; N, 15.92; Cl, 8.47.

5.451-[4-(2,4-Difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-M-isoindol-5-ylmethyl]-urea

A mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.40 g, 1.1 mmol) and CDI (0.18 g, 1.1 mmol)in DMF (5 mL) was stirred at room temperature for 18 h. To the mixturewas added 4-(2,4-difluoro-phenyl)-thiazol-2-ylamine (0.23 g, 1.1 mmol)at room temperature, and the mixture was stirred at 100° C. for 2 days.To the mixture was added water (25 mL) and ethyl acetate (20 mL). Themixture was stirred at room temperature for 2 h. The suspension wasfiltered and the solid was washed with water (20 mL), ethyl acetate (20mL) and water (20 mL) to give a solid. The solid was purified with PrepHPLC to give1-[4-(2,4-difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureaas a white solid (70 mg, 13% yield): HPLC: Waters Symmetry C18, 5 μm,3.9×150 mm, 1 mL/min, 240 nm, 40/60 CH₃CN/0.1% H₃PO₄, 6.43 min (97.7%);mp: 264-266° C.; ¹H NMR (DMSO-d₆) 1.93-2.05 (m, 1H, CHH), 2.30-2.45 (m,1H, CHH), 2.55-2.68 (m, 1H, CHH), 2.82-3.00 (m, 1H, CHH), 4.32 (d,J=17.4 Hz, 1H, CHH), 4.41-4.54 (m, 3H, CH₂, CHH), 5.11 (dd, J=5.1, 13.2Hz, 1H, NCH), 7.09-7.24 (m, 2H, Ar, NH), 7.28-7.41 (m, 2H, Ar), 7.46 (d,J=7.7 Hz, 1H, Ar), 7.53 (s, 1H, Ar), 7.71 (d, J=7.9 Hz, 1H, Ar),7.93-8.10 (m, 1H, Ar), 10.85 (s, 1H, NH), 10.98 (s, 1H, NH); ¹³C NMR(DMSO-d₆) □ 22.48, 31.18, 42.94, 47.12, 51.57, 104.54 (t, JC-F=26.4 Hz),110.67 (d, JC-F=14.3 Hz), 111.81 (dd, JC-F=3.3, 20.9 Hz), 118.91 (dd,JC-F=2.8, 11.6 Hz), 121.95, 123.03, 126.95, 130.29 (dd, JC-F=4.4, 9.9Hz), 130.49, 141.44, 142.45, 143.95, 154.02, 159.48 (dd, JC-F==252.2,12.1 Hz), 159.36, 161.24 (dd, JC-F=246.9, 13.2 Hz), 167.88, 170.98,172.85; LCMS MH=512; Anal. Calcd for C24H19N5O4F2S: C, 56.36; H, 3.74;N, 13.69. Found: C, 56.16; H, 3.80: N, 14.07.

5.466-{3-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester

3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonate (1.11 g, 3.0 mmol) and 1,1′-Carbonyldiimidazole (535mg, 3.3 mmol) were suspended in dry DMF (20 mL) and the mixture wasstirred at rt for 24 h. While stirring, a portion of the reactionmixture (6.7 mL, ˜1 mmol) was transferred to a vial containingtert-butyl 6-amino-3,4-dihydroisoquinoline-2(1H)-carboxylate (273 mg,1.1 mmol). The resulting mixture was stirred at rt overnight and thereaction progress was monitored by LCMS. After 48 h, additionaltert-butyl 6-amino-3,4-dihydroisoquinoline-2(1H)-carboxylate (50 mg, 0.2mmol) was transferred to the reaction mixture and stirring continued foranother 24 h. The reaction mixture was acidified with acidic acid andwater. The volatiles were removed in vacuo and the residue was dissolvedin DMF and purified using C-18 preparatory HPLC to give6-{3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester as a white solid (290 mg, 53% yield): HPLC: WatersSymmetry C₁₈, 5 μm, 3.9×150 mm, 1 ml/min, 240 nm, 40/60 CH₃CN/0.1%H₃PO₄, 5.02 min (96.5%); mp: 230-232° C.; ¹H NMR (DMSO-d₆) δ 1.42 (s,9H, (CH₃)₃), 1.90-2.11 (m, 1H, CHH), 2.23-2.47 (m, 1H, CHH), 2.60 (d,J=17.9 Hz, 1H, CHH), 2.70 (t, J=5.6 Hz, 2H, CH₂), 2.81-3.06 (m, 1H,CHH), 3.51 (t, J=5.7 Hz, 2H, CH₂), 4.18-4.58 (m, 6H, CH₂, CH₂, CH₂),5.11 (dd, J=4.9, 13.2 Hz, 1H, CH), 6.72 (t, J=5.9 Hz, 1H, NH), 7.00 (d,J=8.3 Hz, 1H, Ar), 7.17 (d, J=8.1 Hz, 1H, Ar), 7.27 (br. s., 1H, Ar),7.44 (d, J=7.9 Hz, 1H, Ar), 7.51 (s, 1H, Ar), 7.69 (d, J=7.9 Hz, 1H,Ar), 8.56 (s, 1H, NH), 10.98 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 22.46,28.07, 28.44, 31.16, 41.51, 42.75, 45.04, 47.08, 51.55, 78.81, 116.01,117.49, 121.87, 122.90, 126.20, 126.36, 126.88, 130.25, 134.70, 138.56,142.35, 144.86, 153.97, 155.21, 167.93, 170.96, 172.83; LCMS: MH=548;Anal Calcd for C₂₉H₃₃N₅O₆+1.1 H₂O: C, 61.39; H, 6.25; N, 12.34. Found:C, 61.38; H, 6.11; N, 12.29.

5.47 Isoindolin Compounds of Formula III

The isoindolin compounds of Formula III are made as shown in Scheme 1.

To a solution of the 5-aminomethylisoindolinone glutarimide 1 (1.134mmol) in dry DMF (10 mL) is added Cs₂CO₃ (406 mg, 1.248 mmol). After 5minutes of stirring, an appropriate chloromethoxycarbonyl reagent 2(1.248 mmol) is added and the resulting mixture is stirred at 50° C.overnight. The reaction mixture is neutralized with 0.5 N HCl and thenconcentrated to dryness in vacuo. The residue is purified by preparativeHPLC, followed by the removal of protecting groups, if necessary, understandard conditions to yield a compound of Formula III.

5.48 2,2-Dimethyl-propionic acid3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl}-2,6-dioxo-piperidin-1-ylmethylester

To a solution of1-(3-chloro-4-methyl-phenyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea(500 mg, 1.134 mmol) in dry DMF (10 mL) was added Cs₂CO₃ (406 mg, 1.248mmol). After 5 min of stirring, chloromethyl pivalate (188 mg, 1.248mmol) was added and the resulting mixture was stirred at 50° C.overnight. The reaction mixture was neutralized with 0.5N HCl and thenconcentrated to dryness in vacuo. The residue was dissolved inacetonitrile and water was added slowly resulting in a precipitate toform. The solid was collected by filtration and dissolved in DMF forpurification on a C-18 preparative HPLC column (50/50 CH₃CN/water).Fractions containing the desired product were combined, concentrated invacuo to remove most of the CH₃CN, and then lyophilized to provide theproduct as a white solid (81 mg, 13% yield):HPLC: Waters Symmetry C₁₈, 5μm, 3.9×150 mm, 1 mL/min, 240 nm, 55/45 CH₃CN/0.1% H₃PO₄, 4.58 min(97.2%); mp: 155-157° C.; ¹H NMR (DMSO-d₆) δ 0.95-1.23 (m, 9H, ^(t)Bu),1.96-2.15 (m, 1H, CH), 2.23 (s, 3H, CH₃), 2.31-2.47 (m, 1H, CH), 2.83(d, J=16.6 Hz, 1H, CH), 3.00-3.22 (m, 1H, CH), 4.17-4.60 (m, 4H, CH₂,CH₂), 5.30 (dd, J=5.0, 13.3 Hz, 1H, CH), 5.47-5.83 (m, 2H, CH₂), 6.79(t, J=5.9 Hz, 1H, CH₂NH), 7.02-7.28 (m, 2H, Ar), 7.36-7.60 (m, 2H, Ar),7.60-7.86 (m, 2H, Ar), 8.74 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 18.73,21.56, 26.62, 31.21, 38.21, 42.79, 47.04, 52.01, 63.39, 116.45, 117.66,121.95, 123.01, 126.98, 127.42, 130.11, 131.00, 132.99, 139.58, 142.36,144.87, 155.07, 167.96, 170.13, 171.23, 176.51; LCMS: MH⁺=555, 557;Anal. Calcd. for C₂₈H₃₁ClN₄O₆+0.2 H₂O: C, 60.20; H, 5.67; N, 10.03; Cl,6.35. Found: C, 60.20; H, 5.74; N, 9.94; Cl, 6.39.

5.49(3-(5-((3-(3-Chloro-4-methylphenyl)ureido)methyl)-1-oxoisoindolin-2-yl)-2,6-dioxopiperidin-1-yl)methylpiperidine-4-carboxylate hydrochloride

To 1-tert-butyl4-(3-(5-((3-(3-chloro-4-methylphenyl)ureido)methyl)-1-oxoisoindolin-2-yl)-2,6-dioxopiperidin-1-yl)methylpiperidine-1,4-dicarboxylate (300 mg, 0.44 mmol, obtained as describedin previous example) was added 2 N HCl in Et₂O (15 mL). The slurry wasvigorously stirred for 2 h at rt. The solid was filtered, washed withcopious Et₂O, and dried in a vacuum oven overnight to give(3-(5-(3-(3-chloro-4-methylphenyl)ureido)methyl)-1-oxoisoindolin-2-yl)-2,6-dioxopiperidin-1-yl)methylpiperidine-4-carboxylate Hydrochloride as a white solid (285 mg, 105%yield): HPLC:Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 ml/min, 240 nm,30/70 CH₃CN/0.1% H₃PO₄, 3.26 min (98.2%); mp: 234-236° C.; ¹H NMR(DMSO-d₆) δ 1.62-1.81 (m, 2H, CHH, CHH), 1.87-2.01 (m, 2H, CHH, CHH),2.02-2.16 (m, 1H, CHH), 2.16-2.29 (m, 3H, CH₃), 2.32-2.47 (m, 1H, CHH),2.59-2.76 (m, 1H, CHH), 2.77-2.99 (m, 3H, CHH, CHH, CHH), 3.00-3.28 (m,3H, CHH, CHH, CHH), 4.15-4.66 (m, 4H, CH₂, CH₂), 5.26 (dd, J=4.9, 13.2Hz, 1H, CH), 5.55-5.80 (m, 2H, CH₂O), 7.02 (t, J=5.9 Hz, 1H, NH),7.09-7.29 (m, 2H, Ar), 7.45 (d, J=7.9 Hz, 1H, Ar), 7.53 (s, 1H, Ar),7.62-7.82 (m, 2H, Ar), 8.41-8.96 (m, 2H, NH, NH), 9.09 (s, 1H, NH); ¹³CNMR (DMSO-d₆) δ 18.73, 21.46, 24.31, 31.24, 37.30, 42.00, 42.73, 47.20,52.09, 63.40, 116.31, 117.50, 121.91, 122.99, 126.97, 127.27, 130.09,130.99, 132.96, 139.69, 142.38, 144.94, 155.21, 167.96, 170.19, 171.29,172.20; LCMS: MH=582, 584; Anal Calcd for C₂₉H₃₃C₁₂N₅O₆+1.6H₂O+0.6HCl+0.2 Et₂O+0.4 ^(t)BuCl: C, 53.35; H, 6.05; N, 9.91; Cl, 13.04.Found: C, 53.13; H, 6.06; N, 9.70; Cl, 13.25.

5.50 1-tert-Butyl4-(3-(5-((3-(3-chloro-4-methylphenyl)ureido)methyl)-1-oxoisoindolin-2-yl)-2,6-dioxopiperidin-1-yl)methylpiperidine-1,4-dicarboxylate

Step 1: 1-tert-Butyl 4-chloromethyl piperidine-1,4-dicarboxylate

To a mixture of 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (3g, 13.1 mmol) in dichloromethane (40 mL) and water (50 mL) was addedNaHCO₃ (4.4 g, 52.3 mmol) and tetrabutylammonium hydrogen sulfate (444mg, 1.3 mmol). After stirring the mixture in an ice bath at 0° C. for˜10 min, chloromethyl chlorosulfate (2.59 g, 15.7 mmol) in 10 mL ofdichloromethane was added dropwise. The reaction mixture was allowed towarm up to rt and stirred vigorously overnight. The mixture wastransferred to a separatory funnel with dichloromethane and water (200mL, each). The organic layer was washed with additional water and brine,dried (Na₂SO₄), and concentrated in vacuo to give 1-tert-butyl4-chloromethyl piperidine-1,4-dicarboxylate as a clear oil (3.5 g, 96%yield). This material was used without further purification. ¹H NMR(DMSO-d₆) δ 1.26-1.48 (m, 11H, CHH, CHH, tertBu), 1.83 (dd, J=3.0, 13.2Hz, 2H, CHH, CHH), 2.58-2.75 (m, 1H, CH), 2.74-2.99 (m, 2H, CHH, CHH),3.61-4.02 (m, 2H, CHH, CHH), 5.87 (s, 2H, CH₂O); ¹³C NMR (DMSO-d₆) δ27.16, 27.98, 39.58, 42.35, 69.41, 78.67, 153.75, 172.25; LCMS: MH=278.

Step 2: 1-tert-Butyl 4-iodomethyl piperidine-1,4-dicarboxylate

To a solution of NaI (1.6 g, 11.09 mmol) in dry acetonitrile (8 mL) wasadded dropwise 1-tert-butyl 4-chloromethyl piperidine-1,4-dicarboxylate(2.8 g, 10.08 mmol) in 2 mL of acetonitrile. The mixture turned darkorange and was stirred at rt in the dark for 24 h. The reaction mixturewas filtered to remove NaCl and the filtrate was concentrated in vacuo.The remaining residue was partitioned between DCM and 5% aq NaHSO₃solution. The organic layer was washed with water and brine, dried(Na₂SO₄), and concentrated in vacuo to give 1-tert-butyl 4-iodomethylpiperidine-1,4-dicarboxylate as a pale yellow oil (3.5 g, 94%). Attemptsto characterize by NMR in DMSO-d₆ resulted in rapid decomposition. Thematerial was stored in the dark at −20° C. to minimize decompositionbefore further use. LCMS: MH=370.

Step 3: 1-tert-Butyl4-(3-(5-((3-3-chloro-4-methylphenyl)ureido)methyl)-1-oxoisoindolin-2-yl)-2,6-dioxopiperidin-1-yl)methylpiperidine-1,4-dicarboxylate

1-(3-Chloro-4-methyl-phenyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea(400 mg, 0.907 mmol) and NaH (43.5 mg, 1.82 mmol) were stirred in dryDMF (15 mL) for ˜10 min. To the mixture was added 1-tert-butyl4-iodomethyl piperidine-1,4-dicarboxylate (402 mg, 1.09 mmol) and thereaction stirred in the dark at rt for 2 h. The reaction was quenchedwith acetic acid (5 mL) and concentrated in vacuo. The residue waspartitioned in water and EtOAc (100 mL, each). The separated organiclayer was washed with additional water and brine, dried (Na₂SO₄), andconcentrated in vacuo to give 1.4 gm of a yellowish solid which wasdissolved in DMF for purification on a C-18 prep HPLC column. Fractionscontaining the desired product were combined, concentrated in vacuo toremove most of the CH₁CN, and then lyophilized to provide 1-tert-butyl4-(3-(5-((3-(3-chloro-4-methylphenyl)ureido)methyl)-1-oxoisoindolin-2-yl)-2,6-dioxopiperidin-1-yl)methylpiperidine-1,4-dicarboxylate as a white solid (380 mg, 49% oftheoretical yield from two separate runs that were combined before prepHPLC). HPLC:Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 ml/min 240 nm,55/45 CH₃CN/0.1% H₃PO₄, 5.82 min (99.4%); mp: 157-159° C.; ¹H NMR(DMSO-d₆) δ 1.23-1.51 (m, 11H, (CH₃)₃, CHH, CHH), 1.68-1.84 (m, 2H, CHH,CHH), 1.97-2.13 (m, 1H, CHH), 2.15-2.31 (m, 4H, CH₃, CH), 2.32-2.46 (m,1H, CHH), 2.68-2.94 (m, 3H, CHH, CHH, CHH), 2.98-3.21 (m, 1H, CHH),3.69-3.89 (m, 2H, CHH, CHH), 4.20-4.57 (m, 4H, CH₂, CH₂), 5.27 (dd,J=5.0, 13.3 Hz, 1H, CH), 5.53-5.73 (m, 2H, CH₂O), 6.81 (t, J=5.9 Hz, 1H,NH), 7.06-7.24 (m, 2H, Ar), 7.45 (d, J=7.9 Hz, 1H, Ar), 7.53 (s, 1H,NH), 7.67 (d, J=1.9 Hz, 1H, Ar), 7.71 (d, J=7.7 Hz, 1H, Ar), 8.76 (s,1H, NH); ¹³C NMR (DMSO-d₆) δ 18.66, 21.39, 27.38, 27.97, 31.16, 42.72,47.08, 51.98, 63.15, 78.60, 116.37, 117.60, 121.88, 122.93, 126.91,127.34, 130.06, 130.94, 132.91, 139.52, 142.32, 144.82, 153.74, 155.01,167.90, 170.11, 171.18, 172.97 (two ¹³C signals arising from thepiperidine ring are not observed due to overlap with DMSO-d₆); LCMS:MH=682, 684; Anal Calcd for C₃₄H₄₀ClN₅O₈+1.0 H₂O: C, 58.32; H, 6.05; N,10.00; Cl, 5.06. Found: C, 58.41; H, 5.93; N, 9.96; Cl, 5.22.

5.51N-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-1-imidazol-1-yl-benzamide

A mixture of 4-imidazol-1-yl-benzoic acid (0.21 g, 1.1 mmol) and CDI(0.19 g, 1.2 mmol) in DMF (14 mL) was stirred at 40° C. for 2 hrs. Tothe suspension was added3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.40 g, 1.1 mmol) and the mixture was stirredat 40° C. for 20 hrs. Water (20 mL) was added to the reaction mixtureand the mixture was stirred at RT for 30 min. The suspension wasfiltered and the solid was washed with water (20 mL), ethyl acetate (20mL), and water (20 mL) to give the product as a white solid (340 mg, 71%yield): HPLC: Waters Xterra C₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm, 5%gradient 95% in 5 min, CH₃CN/0.1% NH₄OOCH, 3.92 min (99.0%); mp:268-270° C.; ¹H NMR (DMSO-d₆) δ 1.90-2.08 (m, 1H, CHH), 2.24-2.47 (m,1H, CHH), 2.54-2.67 (m, 1H, CHH), 2.82-3.02 (m, 1H, CHH), 4.31 (d,J=17.4 Hz, 1H, CHH), 4.45 (d, J=17.4 Hz, 1H, CHH), 4.62 (d, J=5.7 Hz,2H, CH₂), 5.11 (dd, J=5.1, 13.2 Hz, 1H, NCH), 7.14 (s, 1H, Ar), 7.49 (d,J=7.9 Hz, 1H, Ar), 7.56 (s, 1H, Ar), 7.71 (d, J=7.7 Hz, 1H, Ar),7.77-7.91 (m, 3H, Ar), 8.05 (d, J=8.7 Hz, 2H, Ar), 8.39 (s, 1H, Ar),9.24 (t, J=5.9 Hz, 1H, NH), 10.98 (br. s., 1H, NH); ¹³C NMR (DMSO-d₆) δ22.49, 31.20, 42.79, 47.13, 51.58, 117.78, 119.59, 122.10, 122.97,127.08, 129.02, 130.20, 130.39, 132.07, 135.60, 138.99, 142.40, 143.83,165.25, 167.92, 170.99, 172.86; LCMS: MH⁺=444; Anal. Calcd. forC₂₄H₂₁N₅O₄+1.5 H₂O: C, 61.27; H, 5.14: N, 14.89. Found: C, 61.03; H,4.93; N, 14.75.

5.52N-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-4H-isoindol-5-ylmethyl]-4-morpholin-4-yl-benzamide

A mixture of 4-morpholin-4-yl-benzoic acid (0.22 g, 1.1 mmol) and CDI(0.19 g, 1.2 mmol) in DMF (4 mL) was stirred at 40° C. for 2 hrs. To thesuspension was added3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.40 g, 1.1 mmol), and the mixture wasstirred at 40° C. for 2 hrs. Water (20 mL) was added to the reactionmixture and the mixture was stirred at RT for 30 min. The suspension wasfiltered and the solid was washed with water (20 mL), ethyl acetate (20mL), and water (20 mL) to give the product as a white solid (370 mg, 74%yield): HPLC: Waters Xterra C₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm, 5%gradient 95% in 5 min, CH₃CN/0.1% NH4OOCH, 1.10 min (96.5%); mp:275-277° C.; ¹H NMR (DMSO-d₆) δ 2.00 (d, J=5.1 Hz, 1H, CHH), 2.38 (m,1H, CHH), 2.54-2.66 (m, 1H, CHH), 2.81-3.01 (m, 1H, CHH), 3.15-3.28 (m,4H, CH₂, CH₂), 3.65-3.80 (m, 4H, CH₂, CH₂), 4.30 (d, J=17.4 Hz, 1H,CHH), 4.44 (d, J=17.6 Hz, 1H, CHH), 4.56 (d, J=5.9 Hz, 2H, CH₂), 5.10(dd, J=5.1, 13.2 Hz, 1H, NCH), 6.98 (d, J=9.1 Hz, 2H, Ar), 7.45 (d,J=7.9 Hz, 1H, Ar), 7.51 (s, 1H, Ar), 7.68 (d, J=7.9 Hz, 1H, Ar), 7.81(d, J=8.9 Hz, 2H, Ar), 8.88 (s, 1H, NH), 10.98 (s, 1H, NH);

¹³C NMR (DMSO-d₆) δ 22.46, 31.15, 42.54, 47.07, 47.33, 51.53, 65.86,113.40, 121.96, 122.86, 123.72, 126.99, 128.50, 130.24, 142.31, 144.38,152.95, 165.87, 167.91, 170.96, 172.83; LCMS: MH⁺=463; Anal. Calcd. forC₂₅H₂₆N₄O₅+0.6 H₂O: C, 63.44; H, 5.79; N, 11.84. Found: C, 63.19; H,5.42; N, 12.09.

5.532-Amino-2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamidehydrochloride

Step 1

To a stirred solution of N-Boc-(4′-chlorophenyl) glycine (1.16 g, 4.10mmol) in DMF (20 mL) was added CDI (0.69 g, 4.30 mmol). The reactionmixture was heated to 40° C. for 2 hrs.3-(5-Aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionehydrochloride (1.50 g, 4.10 mmol) was then added. After 3.5 hrs at 40°C., water (70 mL) was added. Solid precipitated was isolated byfiltration, washed with water (3×20 mL), and dried in vacuo to affordtert-butyl1-(4-chlorophenyl)-2-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methylamino)-2-oxoethylcarbamateas a white solid (1.77 g, 80% yield). The crude product was used in thenext step without further purification.

Step 2

To a stirred solution of tert-butyl1-(4-chlorophenyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methylamino)-2-oxoethylcarbamate(0.84 g, 1.50 mmol) in EtOAc (35 mL) was added a solution of diluted HCl(2N in ether, 30 mL). After 3 days, solid was isolated by filtration,washed with ether (3×10 mL), and dried in vacuo. The solid was thendissolved in water (100 mL) and extracted with EtOAc (2×50 mL). Theaqueous phase was concentrated and the residue was triturated with etherfor 1 hr. The product was isolated by filtration and dried in vacuo togive the product as a white solid (0.65 g, 88% yield): HPLC: WatersSymmetry C₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm, gradient: CH₃CN/0.1%H₃PO₄: 10/90 to 90/10 in 10 min, 90/10 (5 min): 4.91 min (98.00%); mp:255-257° C.; ¹H NMR (DMSO-d₆) δ 1.93-2.09 (m, 1H, CHH), 2.39 (qd, J=4.4,13.1 Hz, 1H, CHH), 2.54-2.69 (m, 1H, CHH), 2.82-3.03 (m, 1H, CHH), 4.24(d, J=17.4 Hz, 1H, CHH), 4.30-4.54 (m, 3H, CH₂, CHH), 5.02-5.19 (m, 2H,CH, CH), 7.22-7.41 (m, 2H, Ar), 7.47-7.71 (m, 5H, Ar), 9.35 (t, J=5.9Hz, 1H, NH), 10.99 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 22.48, 31.20, 42.26,47.03, 51.59, 54.80, 121.89, 121.96, 122.85, 126.97, 128.80, 129.81,130.49, 133.02, 133.99, 142.27, 142.70, 167.22, 167.82, 170.98, 172.85;LCMS: MH⁺=441/443; Anal. Calcd. for C₂₂H₂₂N₄O₄Cl₂+0.6 H₂O: C, 54.13; H,4.79; N, 11.48. Found: C, 53.89; H, 4.82; N, 11.60.

5.542-Amino-5-chloro-N-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-benzamide

DIPEA (1.46 mL, 8.12 mmol) was added to a stirred mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonate (1.0 g, 2.71 mmol) and 5-chloro-isatoic anhydride 3(0.535 g, 2.71 mmol) in acetonitrile (10 mL) under nitrogen. Thereaction mixture was refluxed for 14 hrs, at which time LCMS indicatedthe reaction was complete. Water was added to the slurry and the solidwas collected by filtration, washed with additional water, and dried invacuo to provide the product as a whte solid (860 mg, 74% yield): HPLC:Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm, 30/70CH₃CN/0.1% H₃PO₄, 6.30 min (98.7%); mp: 242-244° C.; ¹H NMR (DMSO-d₆) δ1.79-2.16 (m, 1H, CHH), 2.29-2.46 (m, 1H, CHH), 2.60 (d, J=16.6 Hz, 1H,CHH), 2.74-3.08 (m, 1H, CHH), 4.08-4.69 (m, 4H, CH₂, CH₂), 5.11 (dd,J=4.9, 13.2 Hz, 1H, CH), 6.60 (s, 2H, NH₂), 6.73 (d, J=8.7 Hz, 1H, Ar),7.18 (dd, J=2.4, 8.8 Hz, 1H, Ar), 7.46 (d, J=7.9 Hz, 1H, Ar), 7.53 (s,1H, Ar), 7.59-7.76 (m, 2H, Ar), 9.01 (t, J=5.8 Hz, 1H, NH), 10.98 (s,1H, NH); ¹³C NMR (DMSO-d₆) δ 22.51, 31.20, 42.44, 47.12, 51.56, 114.81,117.71, 118.12, 122.07, 122.91, 127.07, 127.33, 130.35, 131.60, 142.38,143.92, 148.78, 167.67, 167.92, 170.98, 172.85: LCMS: MH⁺=427, 429;Anal. Calcd. for C₂₁H₁₉ClN₄O₄+0.2 H₂O: C, 58.73; H, 4.32; N, 13.05.Found: C, 58.68; H, 4.26; N, 12.80.

5.55 Isoindolin Compounds

The isoindolin compounds shown above are made as shown in Scheme 2.

Step 1

DIPEA (2.0 mmol, 2 eq.) is added to a stirred mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonate 3 (0.26 g, 1.0 mmol) and an appropriatenitrophenylisothiocyanate 4 (1.1 mmol) in acetonitrile (10 mL) undernitrogen. The mixture is stirred at RT for 12 hrs. 1N HCl solution (10mL) is added and the solid formed is filtered, washed with additionalwater, and dried to provide substituted1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(2-nitrophenyl)thiourea5.

Step 2

To a stirred mixture this intermediate 5 (1.0 mmol) in EtOH (10 mL) isadded a solution of sodium dithionite (1.74 g, 10 mmol) in water (10mL). The mixture is heated to 60° C. for 1 hr. The reaction mixture isconcentrated to dryness to yield compound 6, which is used directly inthe next step without further purification.

Step 3

Compound 6 is suspended in DMF and EDCl (0.19 g, 1.0 mmol) is added. Themixture is stirred overnight and the reaction mixture is quenched byaddition of acetic acid (2 mL). The crude product is purified usingpreparative HPLC to provide a compound 7.

5.56 Isoindoline Compounds

According to the procedures provided in Sections 5.34-5.35,1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(2-hydroxy-4,5-dimethylphenyl)ureaare prepared starting from 4,5-dimethyl-2-nitrophenol.

General Procedure A

A mixture of the appropriate carboxylic acid starting material (2.0mmol) and CDI (0.32 g, 2.0 mmol) in DMF (30 mL) is stirred for 2 hoursat 40° C., and then345-(aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dionemethanesulfonate (0.74 g, 2.0 mmol) is added, and stirring proceeds for24 hours. The mixture is evaporated under vacuum and the residue ispurified by preparative HPLC.

Using the General Procedure A, the following compounds are prepared:

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-(morpholinomethyl)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-((4-methylpiperazin-1-yl)methyl)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-methyl-3-(morpholinomethyl)phenyl)acetamide;

2-(4-chloro-3-(morpholinomethyl)phenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-(2-morpholinoethyl)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-methyl-3-(2-morpholinoethyl)phenyl)acetamide;

2-(4-chloro-3-(2-morpholinoethyl)phenyl)-N-((2-(2,6-dioxopiperidin-3)-1-oxoisoindolin-5-yl)methyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-(morpholinomethyl)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-((4-methylpiperazin-1-yl)methyl)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-methyl-4-(morpholinomethyl)phenyl)acetamide;

2-(3-chloro-4-(morpholinomethyl)phenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-(2-morpholinoethyl)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-methyl-4-(2-morpholinoethyl)phenyl)acetamide;

2-(3-chloro-4-(2-morpholinoethyl)phenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-(2-morpholinoethoxy)phenyl)acetamide

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-methyl-4-(2-morpholinoethoxy)phenyl)acetamide

2-(3-chloro-4-(2-morpholinoethoxy)phenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-(2-morpholinoethoxy)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-methyl-3-(2-morpholinoethoxy)phenyl)acetamide;

2-(4-chloro-3-(2-morpholinoethoxy)phenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(morpholinomethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-((4-ethylpiperazin-1-yl)methyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-methyl-4-(morpholinomethyl)benzamide;

3-chloro-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(morpholinomethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(2-morpholinoethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-methyl-4-(2-morpholinoethyl)benzamide;

3-chloro-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(2-morpholinoethyl)benzamide

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(morpholinomethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-((4-methylpiperazin-1-yl)methyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-methyl-3-(morpholinomethyl)benzamide;

4-chloro-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(morpholinomethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(2-morpholinoethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-methyl-3-(2-morpholinoethyl)benzamide;

4-chloro-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(2-morpholinoethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(2-morpholinoethoxy)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-methyl-4-(2-morpholinoethoxy)benzamide;

3-chloro-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(2-morpholinoethoxy)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl-(2-morpholinoethoxy)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-methyl-3-(2-morpholinoethoxy)benzamide;and

4-chloro-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(2-morpholinoethoxy)benzamide5.573-[5-(6-Chloro-4-oxo-4H-quinazolin-3-ylmethyl)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione

2-Amino-5-chloro-N-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-benzamide(250 mg, 0.586 mmol) was suspended in trimethyl orthoformate (3 mL) andthe mixture was heated to 85° C. overnight. Formic acid (100 L) wasadded to the reaction and the mixture was heated for 1 hr. Water wasadded to the reaction resulting in a white precipitate. The solid wasfiltered, washed with additional water, and dried in a vacuum oven toprovide the product as a white solid (140 mg, 55% yield): HPLC: WatersSymmetry C₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm, 30/70 CH₃CN/0.1%H₃PO₄, 7.20 min (97.9%); mp: 308-310° C.; ¹H NMR (DMSO-d₆) δ 1.90-2.08(m, 1H, CHH), 2.37 (qd, J=4.3, 13.2 Hz, 1H, CHH), 2.59 (d, J=17.6 Hz,1H, CHH), 2.78-3.03 (m, 1H, CHH), 4.23-4.35 (m, 1H, NCHH), 4.38-4.51 (m,1H, NCHH), 5.10 (dd, J=5.0, 13.1 Hz, 1H, NCH), 5.33 (s, 2H, NCH₂), 7.52(d, J=7.7 Hz, 1H, Ar), 7.59 (s, 1H, Ar), 7.67-7.80 (m, 2H, Ar), 7.88(dd, J=2.5, 8.7 Hz, 1H, Ar), 8.09 (d, J=2.3 Hz, 1H, Ar), 8.66 (s, 1H,NCHN), 10.98 (s, 1H, CONH); ¹³C NMR (DMSO-d₆) δ 22.45, 31.17, 47.13,49.20, 51.59, 122.68, 122.91, 123.23, 125.11, 127.56, 129.59, 131.14,131.50, 134.59, 140.45, 142.59, 146.68, 148.47, 159.20, 167.63, 170.92,172.82; LCMS: MH⁺=437, 439; Anal. Calcd. for C₂₂H₁₇ClN₄O₄+0.4 H₂O: C,59.51; H, 4.04; N, 12.62; 7.98, Cl. Found: C, 59.29; H, 3.66; N, 12.39;7.82, Cl.

5.586-Chloro-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-1H-Quinazoline-2,4-dione

To a slurry of2-amino-5-chloro-N-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-benzamide(250 mg, 0.586 mmol) in acetonitrile (5 mL) was added DIPEA (0.204 mL,1.171 mmol), followed by phosgene (0.62 mL, 1.171 mmol). The reactionmixture was warmed up to 60° C. After 4 hrs, additional phosgene wasadded (0.31 mL) and stirring was continued for 1 hr, at which time LC-MSindicated reaction completion. The reaction slurry was quenched withdilute HCl, and the white solid was collected by filtration and washedwith additional water and copious Et₂O. The cake was dried in a vacuumoven to afford the product as a white solid (185 mg, 84% yield): HPLC:Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm, 30/70CH₃CN/0.1% H₃PO₄, 7.97 min (97.6%); mp: 358-360° C.; ¹H NMR (DMSO-d₆) δ1.88-2.07 (m, 1H, CHH), 2.24-2.45 (m, 1H, CHH), 2.53-2.68 (m, 1H, CHH),2.79-3.01 (m, 1H, CHH), 4.17-4.36 (m, 1H, CHH), 4.36-4.54 (m, 1H, CHH),5.09 (dd, J=4.9, 13.2 Hz, 1H, CH), 5.19 (s, 2H, PhCH₂), 7.24 (d, J=8.7Hz, 1H, Ar), 7.47 (d, J=7.9 Hz, 1H, Ar), 7.54 (s, 1H, Ar), 7.68 (d,J=7.7 Hz, 1H, Ar), 7.74 (dd, J=2.4, 8.8 Hz, 1H, Ar), 7.89 (d, J=2.3 Hz,1H, Ar), 10.98 (s, 1H, NH), 11.72 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ22.46, 31.18, 43.50, 47.12, 51.58, 115.11, 117.49, 122.29, 122.96,126.37, 126.54, 127.29, 130.65, 135.04, 138.38, 141.17, 142.39, 149.90,161.08, 167.79, 170.94, 172.83. LCMS: MH=453, 455; Anal. Calcd. forC₂₂H₁₇ClN₄O₅+0.3 H₂O: C, 57.66; H, 3.87; N, 12.23; Cl, 7.74. Found: C,57.60; H, 3.90; N, 11.97; Cl, 7.72.

5.59[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-4H-isoindol-5-ylmethyl]-carbamicacid 4-chloro-3-methyl-phenyl ester

To the solution of para-nitro-phenyl chloroformate (1000 mg, 0.5 mmol)in CH₃CN (5 mL), was added the CH₃CN solution (5 mL) of4-chloro-m-cresol (71.3 mg, 0.5 mmol) and DIPEA (0.24 mL, 1.5 mmol)dropwise at room temperature. The mixture was stirred at roomtemperature for 10 min.3-(5-Aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonic acid salt (0.37 g, 1 mmol) was added followed byadditional DIPEA (0.24 mL, 1.5 mmol). The mixture was stirred overnightat ambient temperature. The mixture was then filtered. The resultedsolid was purified on silica gel column eluted using methanol andmethylene chloride to give[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-carbamicacid 4-chloro-3-methyl-phenyl ester as a white solid (30 mg, 14% yield).HPLC: Waters Symmetry C₁₈, 5 μm, 3.9×150 mm, 1 mL/min, 240 nm, 50/50,CH₃CN/0.1% H₃PO₄,: t_(R)=3.0 min (97%); mp 225-227° C.; ¹H NMR (DMSO-d₆)δ 1.91-2.08 (m, 1H, CHH), 2.32 (s, 3H, CH₃), 2.40 (dd, J=4.3, 12.8 Hz,1H, CHH), 2.63 (br. s., 1H, CHH), 2.83-3.02 (m, 1H, CHH), 4.22-4.56 (m,4H, CH₂, CH₂), 5.12 (dd, J=5.1, 13.2 Hz, 1H, NCH), 7.01 (dd, J=2.3, 8.7Hz, 1H, Ar), 7.17 (d, J=2.3 Hz, 1H, Ar), 7.40 (d, J=8.7 Hz, 1H, Ar),7.46 (d, J=7.4 Hz, 1H, Ar), 7.55 (s, 1H, Ar), 7.72 (d, J=7.9 Hz, 1H,Ar), 8.47 (t, J=6.0 Hz, 1H, NH), 10.99 (s, 1H, NH). ¹³C NMR (DMSO-d₆) δ19.52, 22.49, 31.20, 44.04, 47.15, 51.59, 121.03, 122.04, 123.04,124.43, 127.01, 129.33, 130.58, 136.54, 142.45, 143.35, 145.35, 149.62,154.43, 167.87, 170.99, 172.86; LC-MS: 442; Anal Calcd forC₂₂H₂₀ClN₃O₆C, 59.80; H, 4.56; N, 9.51. Found: C, 59.74; H, 4.45; N,9.58.

5.601-[1-(3,4-Dichloro-phenyl)-ethyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea

A stirred mixture of3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethane sulfonate (0.37 g, 1.00 mmol) and CDI (0.18 g, 1.10 mmol) inN,N-dimethylformamide (10 mL) was heated to 40° C. under nitrogenovernight. 1-(3,4-Dichloro-phenyl)-ethylamine (0.19 g, 1.00 mmol) wasthen added and the mixture was heated at 40° C. overnight. Water (60 mL)was added, solid precipitated, was filtered, washed with water (30 mL)and was purified by preparative HPLC (gradient: CH₃CN/H₂O: 10/90 for 5min, to 100/0 in 10 min, 100/0 for 5 min). Solvent was evaporated andthe residue was triturated in ether (20 mL) for 1 h. The product wasthen isolated by filtration and dried in vacuo to give1-[1-(3,4-Dichloro-phenyl)-ethyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureaas a white solid (0.084 g, 17% yield): HPLC: Waters Symmetry C₁₈, 5 μm,3.9×150 mm, 1 mL/min, 240 nm, gradient: CH₃CN/0.1% H₃PO₄: 10/90 to 90/10in 10 min. 90/10 (5 min): 8.57 min (96.68%); mp: 200-202° C.; ¹H NMR(DMSO-d₆) δ 1.32 (d, J=7.0 Hz, 3H, CH₃), 1.88-2.13 (m, 1H, CHH),2.28-2.47 (m, 1H, CHH), 2.55-2.70 (m, 1H, CHH), 2.79-3.04 (m, 1H, CHH),4.18-4.55 (m, 4H, CH₂, CH₂), 4.74 (quin, J=7.0 Hz, 1H, CH), 5.11 (dd,J=4.9, 13.2 Hz, 1H, CH), 6.51 (s, 1H, NH), 6.66 (d, J=7.9 Hz, 1H, NH),7.20-7.46 (m, 3H, Ar), 7.48-7.62 (m, 2H, Ar), 7.65 (d, J=7.7 Hz, 1H.Ar), 10.99 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 22.51, 22.80, 31.20, 42.86,47.07, 48.18, 51.55, 121.61, 122.81, 126.28, 126.70, 127.78, 128.79,130.14, 130.38, 130.77, 142.26, 145.29, 147.41, 157.20, 167.95, 170.98,172.85; LCMS: MH=489, 491; Anal Calcd for C₂₃H₂₂N₄O₄Cl₂+0.9 H₂O: C,54.64, H, 4.75; N, 11.08. Found: C, 54.27, H, 4.66; N, 10.98.

5.611-(3-Chloro-4-(2-(2-methoxyethoxy)ethoxy)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea

3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dionemethanesulfonate (1.11 g, 3.0 mmol) and 1,1′-Carbonyldiimidazole (535mg, 3.3 mmol) were suspended in dry DMF (20 mL) and the mixture wasstirred at rt for 24 h. While stirring, a portion of the reactionmixture (6.7 mL, ˜1 mmol) was transferred to a vial containing3-chloro-4-(2-(2-methoxyethoxy)ethoxy)aniline hydrochloride (310 mg, 1.1mmol). The resulting mixture was stirred at rt overnight and thereaction progress was monitored by LCMS. After 48 h, additional3-chloro-4-(2-(2-methoxyethoxy)ethoxy)aniline hydrochloride (56 mg, 0.2mmol) was transferred to the reaction mixture and stirring continued foranother 24 h. The reaction mixture was acidified with 1 N HCl and waterwas added with stirring until a precipitate formed. The solid wascollected by filtration, suction dried, then dissolved in DMF andpurified using C-18 preparatory HPLC to give1-(3-chloro-4-(2-(2-methoxyethoxy)ethoxy)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureaas a white solid (390 mg, 72% yield): HPLC:Waters Symmetry C₁₈, 5 μm,3.9×150 mm, 1 ml/min, 240 nm, 30/70 CH₁CN/0.1% H₃PO₄, 5.40 min (99.8%);mp: 188-190° C.; ¹H NMR (DMSO-d₆) δ 1.92-2.10 (m, 1H, CHH), 2.29-2.47(m, 1H, CHH), 2.60 (dd, J=2.2, 15.4 Hz, 1H, CHH), 2.82-3.01 (m, 1H,CHH), 3.25 (s, 3H, CH₃), 3.46 (dd, J=3.7, 5.6 Hz, 2H, CH₂), 3.61 (dd,J=3.8, 5.7 Hz, 2H, CH₂), 3.73 (d, J=4.7 Hz, 2H, CH₂), 4.09 (t, J=4.9 Hz,2H, CH₂), 4.24-4.55 (m, 4H, CH₂, CH₂), 5.11 (dd, J=5.1, 13.2 Hz, 1H,CH), 6.77 (t, J=5.9 Hz, 1H, NH), 7.04 (d, J=9.1 Hz, 1H. Ar), 7.19 (dd,J=2.5, 9.0 Hz, 1H, Ar), 7.38-7.49 (m, 1H, Ar), 7.51 (s, 1H, Ar), 7.64(d, J=2.6 Hz, 1H. Ar), 7.69 (d, J=7.7 Hz, 1H, Ar), 8.65 (s, 1H, NH),10.98 (s, 1H, NH); ¹³C NMR (DMSO-d₆) δ 22.51, 31.20, 42.80, 47.12,51.56, 58.05, 68.80, 68.85, 69.84, 71.29, 114.66, 117.58, 119.49,121.24, 121.88, 122.91, 126.89, 130.29, 134.53, 142.38, 144.84, 148.40,155.25, 167.95, 171.01, 172.86; LCMS: MH=545, 547; Anal Calcd forC₂₆H₂₉ClN₄O₇+0.4 H₂O: C, 56.55; H, 5.44; N, 10.15. Found: C, 56.64; H,5.34; N, 10.15.

5.62 Assays

5.62.1 TNFα Inhibition Assay in PMBC

Peripheral blood mononuclear cells (PBMC) from normal donors areobtained by Ficoll Hypaque (Pharmacia, Piscataway, N.J., USA) densitycentrifugation. Cells are cultured in RP 1640 (Life Technologies, GrandIsland, N.Y., USA) supplemented with 10% AB+human serum (GeminiBio-products, Woodland, Calif., USA), 2 mM L-glutamine, 100 U/mlpenicillin, and 100 μg/mL streptomycin (Life Technologies).

PBMC (2×10⁵ cells) are plated in 96-well flat-bottom Costar tissueculture plates (Corning, N.Y., USA) in triplicate. Cells are stimulatedwith LPS (from Salmonella abortus equi, Sigma cat. no. L-1887, St.Louis, Mo., USA) at 1 ng/mL final in the absence or presence ofcompounds. Compounds provided herein are dissolved in DMSO (Sigma) andfurther dilutions are done in culture medium immediately before use. Thefinal DMSO concentration in all assays can be about 0.25%. Compounds areadded to cells 1 hr before LPS stimulation. Cells are then incubated for18-20 hrs at 37° C. in 5% CO₂, and supernatants are then collected,diluted with culture medium and assayed for TNFα levels by ELISA(Endogen, Boston, Mass., USA). IC₅₀s are calculated using non-linearregression, sigmoidal dose-response, constraining the top to 100% andbottom to 0%, allowing variable slope (GraphPad Prism v3.02).

5.62.2 IL-2 and MIP-3a Production by T Cells

PBMC are depleted of adherent monocytes by placing 1×10⁸ PBMC in 10 mLcomplete medium (RPMI 1640 supplemented with 10% heat-inactivated fetalbovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mLstreptomycin) per 10 cm tissue culture dish, in 37° C., 5% CO₂ incubatorfor 30-60 min. The dish is rinsed with medium to remove all non-adherentPBMC. T cells are purified by negative selection using the followingantibody (Pharmingen) and Dynabead (Dynal) mixture for every 1×10⁸non-adherent PBMC: 0.3 mL Sheep anti-mouse IgG beads, 15 μL, anti-CD16,15 μL anti-CD33, 15 μL anti-CD56, 0.23 mL anti-CD19 beads, 0.23 mLanti-HLA class II beads, and 56 pt anti-CD 14 beads. The cells andbead/antibody mixture is rotated end-over-end for 30-60 min at 4° C.Purified T cells are removed from beads using a Dynal magnet. Typicalyield is about 50% T cells, 87-95% CD3⁺ by flow cytometry.

Tissue culture 96-well flat-bottom plates are coated with anti-CD3antibody OKT3 at 5 μg/mL in PBS, 100 μL per well, incubated at 37° C.for 3-6 hrs, then washed four times with complete medium 100 μL/welljust before T cells are added. Compounds are diluted to 20 times offinal in a round bottom tissue culture 96-well plate. Finalconcentrations are about 10 μM to about 0.00064 μM. A 10 mM stock ofcompounds provided herein is diluted 1:50 in complete for the first 20×dilution of 200 μM in 2% DMSO and serially diluted 1:5 into 2% DMSO.Each compound is added at 10 μL per 200 μL culture, to give a final DMSOconcentration of 0.1%. Cultures are incubated at 37° C., 5% CO, for 2-3days, and supernatants analyzed for IL-2 and MIP-3a by ELISA (R&DSystems). IL-2 and MIP-3a levels are normalized to the amount producedin the presence of an amount of a compound provided herein, and EC₅₀sare calculated using non-linear regression, sigmoidal dose-response,constraining the top to 100% and bottom to 0%, allowing variable slope(GraphPad Prism v3.02).

5.62.3 Cell Proliferation Assay

Cell lines Namalwa, MUTZ-5, and UT-7 are obtained from the DeutscheSammlung von Mikroorganismen and Zellkulturen GmbH (Braunschweig,Germany). The cell line KG-1 is obtained from the American Type CultureCollection (Manassas, Va., USA). Cell proliferation as indicated by³H-thymidine incorporation is measured in all cell lines as follows.

Cells are plated in 96-well plates at 6,000 cells per well in media. Thecells are pre-treated with compounds at about 100, 10, 1, 0.1, 0.01,0.001, 0.0001 and 0 μM in a final concentration of about 0.25% DMSO intriplicate at 37° C. in a humidified incubator at 5% CO, for 72 hrs. Onemicrocurie of (Amersham) is then added to each well, and cells areincubated again at 37° C. in a humidified incubator at 5% CO₂ for 6 hrs.The cells are harvested onto UniFilter GF/C filter plates (Perkin Elmer)using a cell harvester (Tomtec), and the plates are allowed to dryovernight. Microscint 20 (Packard) (25 μL/well) is added, and plates areanalyzed in TopCount NXT (Packard). Each well is counted for one min.Percent inhibition of cell proliferation is calculated by averaging alltriplicates and normalizing to the DMSO control (0% inhibition). Eachcompound is tested in each cell line in three separate experiments.Final IC₅₀s are calculated using non-linear regression, sigmoidaldose-response, constraining the top to 100% and bottom to 0%, allowingvariable slope. (GraphPad Prism v3.02).

5.62.4 Immunoprecipitation and Immunoblot

Namalwa cells are treated with DMSO or an amount of a compound providedherein for 1 hr, then stimulated with 10 U/mL of Epo (R&D Systems) for30 min. Cell lysates are prepared and either immunoprecipitated with Eporeceptor Ab or separated immediately by SDS-PAGE. Immunoblots are probedwith Akt, phospho-Akt (Ser473 or Thr308), phospho-Gab1 (Y627), Gab1,IRS2, actin, and IRF-1 Abs and analyzed on a Storm 860 Imager usingImageQuant software (Molecular Dynamics).

5.62.5 Cell Cycle Analysis

Cells are treated with DMSO or an amount of a compound provided hereinovernight. Propidium iodide staining for cell cycle is performed usingCycleTEST PLUS (Becton Dickinson) according to manufacturer's protocol.Following staining, cells are analyzed by a FACSCalibur flow cytometerusing ModFit LT software (Becton Dickinson).

5.62.6 Apoptosis Analysis

Cells are treated with DMSO or an amount of a compound provided hereinat various time points, then washed with annexin-V wash buffer (BDBiosciences). Cells are incubated with annexin-V binding protein andpropidium iodide (BD Biosciences) for 10 min. Samples are analyzed usingflow cytometry.

5.62.7 Luciferase Assay

Namalwa cells are transfected with 4 μg of API-luciferase (Stratagene)per 1×10⁶ cells and 3 μL Lipofectamine 2,000 (Invitrogen) reagentaccording to manufacturer's instructions. Six hours post-transfection,cells are treated with DMSO or an amount of a compound provided herein.Luciferase activity is assayed using luciferase lysis buffer andsubstrate (Promega) and measured using a luminometer (Turner Designs).

5.62.8 Anti-Proliferation Assays

Day 1: The cells are seeded to 96-well plate with 50 ul/well in 10% FBSRPMI (w/Glutamine, w/o pen-strip) for overnight. The following cells areused:

Colorectal cancer cell: Colo 205 3200 cells/well: positive controlirinotecan

Pancreatic cancer cell: BXPC-3 1200 cells/well; positive controlgemcitabine

Prostate cancer cell: PC3 1200 cells/well; positive control docetaxel

Breast cancer cell: MDA-MB-231 2400 cells/well; positive controlpaclitaxel

Day 2: The compounds are serially diluted from 0.00001 μm˜10 μm (or0.000001˜1 μM) with 50 μl/well (of 2×) and added to the plates induplicate with relative positive control. The plates were then incubatedat 37° C. for 72 hours.

Day 5: The results are detected by CellTiter Glo method. 100 μl/well ofCellTiter Glo reagent is added to the plates and incubated for 10minutes at room temperature, and then analyzed on the Top Count reader.The IC₅₀ of each compound is typically based on the result of two ormore individually experiments.

5.63 TNFα Inhibition

The properties of certain compounds provided herein in inhibiting TNFαwere assessed using procedures substantially similar to those describedin Section 6.62.1 above. Tested compounds included:1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-hydroxymethyl-phenyl)-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-imidazol-1-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[4-(4-methyl-4H-[1,2,4]triazol-3-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea;1-(3-amino-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea;N-(4-{3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]ureido}-phenyl)-acetamide;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea;1-[3-(1H-benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-(4-tert-butyl-cyclohexyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea;1-(6-chloro-pyridin-3-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-[4-(2,4-difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;and 2,2-dimethyl-propionic acid3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl}-2,6-dioxo-piperidin-1-ylmethylester. The IC₅₀ values for all of the tested compounds were determinedto be in the range of about 0.2 to 300 nM.

5.64 Antiproliferation—Namalwa

The antiproliferation of certain compounds provided herein were assessedusing Namalwa cells by following procedures substantially similar tothose described in Section 5.62.3 above. The tested compounds included:1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-hydroxymethyl-phenyl)-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-imidazol-1-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea;1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea;1-(3-amino-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea;1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(3-methyl-[1,2,4]oxadiazol-5-yl)-phenyl]-urea;1-(4-tert-butyl-cyclohexyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea;and 2,2-dimethyl-propionic acid3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl}-2,6-dioxo-piperidin-1-ylmethylester. The IC₅₀ values for all of the tested compounds were determinedto be in the range of about 0.02 to 40 nM.

5.65 Antiproliferation—PC3

The antiproliferation of certain compounds provided herein were assessedusing PC3 cells by following procedures substantially similar to thosedescribed in Section 5.62.8 above. The tested compounds included:1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea;1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea:1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea;1-(3-amino-4-methylphenyl)-3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea;1-[3-(1H-benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-(4-tert-butyl-cyclohexyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea;1-(6-chloro-pyridin-3-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-[4-(2,4-difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;2,2-dimethyl-propionic acid3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl}-2,6-dioxo-piperidin-1-ylmethylester; and1-[1-(3,4-dichloro-phenyl)-ethyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea.The IC₅₀ values for all of the tested compounds were determined to be inthe range of about 0.001 to 0.74 μM.

5.66 Antiproliferation—BxPC3

The antiproliferation of certain compounds provided herein were assessedusing BxPC3 cells by following procedures substantially similar to thosedescribed in Section 5.62.8 above. The tested compounds included:1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea;1-(3-amino-4-methylphenyl)-3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea;1-[3-(1H-benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-(4-tert-butyl-cyclohexyl)-342-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl)urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea;1-[4-(2,4-difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;2,2-dimethyl-propionic acid3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl}-2,6-dioxo-piperidin-1-ylmethylester; and1-[1-(3,4-dichloro-phenyl)-ethyl]-3,2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl)urea.The IC₅₀ values for all of the tested compounds were determined to be inthe range of about 0.01 to 0.94 μM.

5.67 Antiproliferation—MDAMB321

The antiproliferation of certain compounds provided herein were assessedusing MDAMB321 cells by following procedures substantially similar tothose described in Section 5.62.8 above. The tested compounds included:1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-1-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea;1-(3-amino-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea;1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(3-methyl-[1,2,4]oxadiazol-5-yl)-phenyl]-urea;1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea;1-[3-(1H-benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-(4-tert-butyl-cyclohexyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea;1-(6-chloro-pyridin-3-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-[4-(2,4-difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;2,2-dimethyl-propionic acid3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl}-2,6-dioxo-piperidin-1-ylmethylester; and1-[1-(3,4-dichloro-phenyl)-ethyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea.The IC₅₀ values for all of the tested compounds were determined to be inthe range of about 0.4 to 380 nM.

5.68 Antiproliferation—Colo205

The antiproliferation of certain compounds provided herein were assessedusing Colo205 cells by following procedures substantially similar tothose described in Section 5.62.8 above. The tested compounds included:1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea:1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea;1-(3-amino-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea;1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea;1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(3-methyl-[1,2,4]oxadiazol-5-yl)-phenyl]-urea:1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea;1-[3-(1H-benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-(4-tert-butyl-cyclohexyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea;1-(6-chloro-pyridin-3-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;1-[4-(2,4-difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]urea;2,2-dimethyl-propionic acid3-[5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl]-2,6-dioxo-piperidin-1-ylmethylester; and1-[1-(3,4-dichloro-phenyl)-ethyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea.The IC₅₀ values for all of the tested compounds were determined to be inthe range of about 0.15 to 130 nM.

The examples set forth above are provided to give those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the claimed embodiments, and are not intended to limit thescope of what is disclosed herein. Modifications that are obvious topersons of skill in the art are intended to be within the scope of thefollowing claims. All publications, patents, and patent applicationscited in this specification are incorporated herein by reference intheir entireties as if each such publication, patent or patentapplication were specifically and individually indicated to beincorporated herein by reference.

1. A compound of Formula V:

or a pharmaceutically acceptable salt, prodrug, or stereoisomer thereof,wherein: X is C(═O) or CH₂; m is an integer of 0, 1, 2, or 3; R¹⁰ andR¹¹ are each independently hydrogen, halo, C₁₋₆ alkyl, or C₆₋₁₀ aryloxy,wherein the alkyl and aryl are each optionally substituted with one ormore halo.
 2. The compound of claim 1, which is:

or a pharmaceutically acceptable salt, prodrug, or stereoisomer thereof.3. A pharmaceutical composition comprising the compound of claim 1, andone or more pharmaceutically acceptable excipients or carriers.
 4. Thepharmaceutical composition of claim 3, further comprising a secondtherapeutic agent.
 5. The pharmaceutical composition of claim 3, whereinthe composition is formulated for single dose administration.
 6. Thepharmaceutical composition of claim 3, wherein the composition isformulated as oral, parenteral, or intravenous dosage form.
 7. Thepharmaceutical composition of claim 6, wherein the oral dosage form is atablet or capsule.
 8. A method of treating or managing cancer comprisingadministering to a subject a therapeutically effective amount of acompound of claim 1, wherein the cancer is Non-Hodgkin's lymphoma,Hodgkin's lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma,or cutaneous T-cell lymphoma.
 9. The method of claim 8, which furthercomprises administration of one or more additional agents.
 10. Themethod of claim 8, wherein the compound is administered orally orparenterally.
 11. A method of treating or managing cancer comprisingadministering to a subject a therapeutically effective amount of acompound of claim 1, wherein the cancer is prostate cancer, pancreaticcancer, breast cancer or colorectal cancer.
 12. The method of claim 11,which further comprises administration of one or more additional agents.13. The method of claim 11, wherein the compound is administered orallyor parenterally.
 14. The compound of claim 1, wherein X is CH₂.
 15. Thecompound of claim 1, wherein m is 1.